MXPA00011567A - Poly(adp-ribose)polymerase gene - Google Patents

Abstract

The invention relates to poly(ADP-ribose)polymerase (PARP) homologues which are characterised by an amino acid sequence with a) a functional NAD+-binding site and b) no zinc-finger-sequence motif of general formula CX2CXmHX2C, wherein m is an integral number 28 or 30 and the radicals X represent any amino acid, independently of each other;and to the functional equivalents of said poly(ADP-ribose)polymerase (PARP) homologues. The invention also relates to nucleic acids coding the poly(ADP-ribose)polymerase (PARP) homologues, to antibodies with specificity for the novel protein, to pharmaceutical and gene therapy agents containing the inventive products, to methods for analytically determining the inventive proteins and nucleic acids, to methods for identifying the effectors or bonding partners of the inventive proteins, to novel PARP effectors and to methods for determining the effectiveness of effectors of this type.

Description

NOVEDOUS GENES OF (ADP-RIBOSE) POLYMERASE The present invention relates to novel poly (ADP-ribose) polymerase (PARP) genes and to proteins derived therefrom; to antibodies with specificity for novel proteins; to pharmaceutical and gene therapy compositions comprising products according to the invention; to methods for the analytical determination of proteins and nucleic acids according to the invention; to methods for identifying effectors or base pairs of the proteins according to the invention; to methods to determine the activity of said effectors and to the use thereof for the diagnosis or therapy of pathological conditions. In 1966, Chambón et al. Discovered a 116 kD enzyme that was characterized in detail in subsequent years and is currently called PARP (EC 2.4.2.30) (poly (adenosine-5'-diphosphoribosa) polymerase), PARS (poly (adenosine) 5 '-difosfo-rribose) synthetase) or ADPRT (adenosine-5' -difosforribose transferase). In the plant kingdom (Arabidopsis thaliana) a PARP of 72 kD (637 amino acids) was discovered in 1995 (Lepiniec L. et al., FEBS Lett 1995; 364 (2): 103-8). It was not clear if this shorter form of PARP is a plant-specific individuality or an artifact (splice variant or similar). To date, the 116 kD PARP enzyme has been unique in animals and humans for its activity, which is described below. It is called PARP 1 to "avoid ambiguities.

The primary physiological function of PARP 1 appears to be its participation in a complex repair mechanism that cells have developed to repair broken DNA strands. The primary cellular response to a strand break of DNA seems to consist rather in the synthesis, catalyzed by PARP 1, of 'poly (ADP-ribose) from NADT (compare with De Murcia, G. et al (1994 ) TIBS, 19, 172). PARP 1 has a modular molecular structure. Three functional elements have been identified to date major: an N-terminal domain for binding to 46 kD DNA; a 22 kD self-modifying central domain to which poly (ADP-ribose) binds, decreasing the enzymatic activity of PARP 1 with increasing elongation; and a C-terminal domain for NAD + binding of 54 kD. It has been found a leucine zipper region within the self-modifying domain, indicating possible protein-protein interactions, only in the PARP from Drosophila. All PARPs known to date are presumably active as homodimers. 20 The high degree of organization of the molecule is reflected in the strong conservation of the amino acid sequence. Thus, 62% conservation of the amino acid sequence for PARP 1 of humans, mice, cattle and chickens has been found. There are • major structural differences with regarding the PARP of Drosophila. The domains themselves ^^ i ^^^^^ É ^^. ^^^^^^^^^^^ gi ^^^^^ ga ^^^ fa ^^^^^^ il ^^^ ¡i8s¡ ^^^ ^^^^^^ - ^^^^^^^ h ~ ^ fi '' * '* * individuals have, in turn, massifs of increased conservation. Thus, the DNA binding region contains two subdomains called zinc fingers (comprising patterns of type CX2CX2s / 3oHX; C), which are involved in Zn2 + recognition -dependent of breaks in DNA monoheads or single-stranded DNA (by eg at the ends of the chromosomes, the telomeres). The catalytic C-terminal domain comprises a block of approximately 50 amino acids (residues 859-908), which is approximately 100% conserved among vertebrates ("signature" of PARP) .This block binds to the natural substrate NADT and thus governs the synthesis of poly (ADP-ribose) (compare Murcia, loe. cit.) The GX3GKG pattern in particular is characteristic of PARPs in this block.The beneficial function described above contrasts with a pathological one in numerous diseases (stroke, myocardial infarction, sepsis, etc.). PARP is involved in cell death resulting from cerebral ischemia (Choi, D., (1997) Nature Medicine, 3, 10, 1073), from the myocardium (Zingarelli, B. , et al (1997), Cardiovascular Research, 36, 205) and ocular (Lam, TT (1997), Res. Co m.in Molecular Pathology and Pharmacology, 95, 3, 241.) Activation of PARP induced by Inflammatory mediators have also been observed in septic shock (Szabo, C, et al. (1997), Jo urnal of Clinical Investigation, 100, 3, 723).

?? In these cases, the activation of the PARP is accompanied by an extensive NAD + consumption. Since four moles of ATP are consumed in the biosynthesis of one mole of NAD +, the reserve of cellular energy decreases drastically. The consequence is cell death. The PARP inhibitors described in the aforementioned specialized literature are nicotinamide and 3-aminobenzamide. 3, 4-Dihydro-5- [4- (1-piperidinyl) butoxy-1 (2H) -isoquinolone is described by Takahashi, K., et al. (1997), Journal of Cerebral Blood Flow and Metabolism 17, 1137. Other inhibitors are described, for example, in Banasik, M., et al. (1992) J. Biol. Chem., 267, 3, 1569 and Gnffin, R.J., et al. (1995), Anti-Cancer Drug Design, 10, 507. The b.p. High molecular weight proteins described for human PARP 1 include the base cleavage repair protein (BER) XRCC1 (X-ray repair cross-complement 1), which is attached by means of a zinc finger pattern, and a module (amino acids 372-524) of BRCT (BRCA1, C-terminal end) (Masson, M., et al., (1998) Molecular and Cellular Biology, 18, 6, 3563). It is an objective of the present invention, due to the various physiological and pathological functions of PARP, to provide novel PARP homologs. The reason is that the provision of homologous PARPs would be particularly important to develop novel targets for drugs, and novel drugs, in order to improve the diagnosis and / or therapy of pathological conditions in which PARP, PARP homologs, are involved. substances derived from them. We have discovered that this objective is achieved by providing PARP homologs, preferably derived from human mammals and human np, possessing an amino acid sequence having: a) a functional domain of NADT binding, ie a "signature" sequence of PARP that have the characteristic pattern GX3GKG, and b) especially in the region of the N-terminal sequence, that is in the region of the first 200, as, for example, in the region of the first 100, N-terminal amino acids, no pattern of PARP zinc finger type sequence of general formula CX-CXr iX? C in which m is an integer between 28 and 30, and the X radicals are, independently of each other, any amino acids, and the functional equivalents thereof Since the PARP molecules according to the present invention represent functional homologues in particular, they naturally also have poly (ADP-ribose) synthesis activity. it corresponds # t * • £. essentially to this activity and it is located towards the C terminal end. Thus, an essential characteristic of the PARPs according to the invention is the presence of a functional NAD binding domain "(signature of PARP) that is located in the C-terminal region of the amino acid sequence (ie approximately in the region of the last 400, such as, for example, the last 350 or 300 C-terminal amino acids), in combination with an N-terminal sequence that has no zinc finger pattern, such as zinc finger patterns in known PARPs presumably contribute to the recognition of DNA breaks, it is assumed that the proteins according to the invention do not interact with the DNA or do otherwise.It has been demonstrated by suitable biochemical assays that the PARP2 according to the invention can be activated by "activated DNA" (ie the DNA after a limited digestion with DNasal) It can also be concluded from this that the PARP2 according to the invention has DN binding properties. A. However, the mechanism of DNA binding and enzymatic activation differs between the PARPs according to the invention and PARP1. As mentioned, its binding to DNA and its enzymatic activation are mediated by a characteristic pattern of zinc finger. There are no such patterns present in the PARPs according to the invention. Presumably these properties are mediated by positively charged amino acids from the N-terminal region of the PARPs according to the invention. Since the 'activated DNA' (ie for example a DNA after a limited treatment with Dnasal) has a large number of defects (interruptions of a single strand, single-strand gaps, single-strand leftovers, double-strand interruptions, etc.), it is possible that although PARP1 and PARPs according to the present invention are activated by the same 'activated DNA', either through of a different subset of defects (eg single-strand breaks instead of single-strand breaks). The functional NAD-binding domain "(ie the catalytic domain) binds to the substrate for the synthesis of poly (ADP-ribose). Consistent with the known PARPs, the sequential pattern GX "X-? JGKG, in which G is glycine, K is lysine, and X :, X" and X3 are, independently of each other, any amino acids, is present in particular . However, as surprisingly demonstrated by comparison of the amino acid sequences of the NAD + binding domains of the PARP molecules according to the invention with the human PARP1 previously described, the sequences according to the invention differ markedly from the sequence known for the NAD + binding domain. A group of preferred PARP molecules according to the invention preferably have in common the following general sequential pattern in the catalytic domain: PXn (S / T) GX? GKGIYFA (SEQ ID NO: 11), in particular (S / T) XGLR (I / V) XPXn (S / T) GX3GKGIYFA (SEQ ID NO: 12), preferably LLWHG (S / T) X-IL (S / T) XGLR (I / V) XPXr? (S / T) GX; GKGIYFAX;, SKSAXY (SEQ ID NO: 13) in which (S / T) describes the alternating occupation of this position in the sequence by S or T, (I / V) describes the alternating occupation of this position in the sequence by I or V, and n is an integer between 1 and 5, and the radi- ers X are, independently of one another, amino acids next. The last pattern is also referred to as the "PARP signature." Similarly, the self-modifying domain is preferably present in the PARPs according to the invention, it may be located, for example, in the region of approximately 100 amino acid. at 200, opposite the N-terminus of the NAD binding domain.The PARP homologs according to the invention may additionally comprise the N-terminal side of the NAD binding domain "(ie between about 30 and about 80 amino acids closer to the N-terminus), a sequential leucine zipper pattern of the general formula: (L / V) X6LX6LX6L (SEQ ID NO: 14) "Jr .., i i in which (L / V) represents the alternating occupation of this sequential position by L or V, and the X radicals are, independently of one another, amino acids that follow. The leucine zipper patterns observed according to the invention differ markedly in position from those described for the Drpsophila PARP. Leucine zipper patterns can lead to homodimers (two molecules of PARP) or heterodimers (a molecule of PARP with an associated molecule that differs from it). The PARP homologs according to the invention preferably additionally comprise the N-terminal side of the leucine zipper patterns, ie between about 10 and about 250 amino acids closer to the N-terminus, at least one more the following partial sequence patterns: LX? NX: YX2QLLX (D / E) Xb GRVG, (pattern 1; SEQ ID NO: 15) AX5EXKX.1KTXNXWX5FX3PXK, (pattern 2; SEQ ID NO: 16) QXL (I / L) XCIX ? MX10PLGKLX; QIX6L, (pattern 3; SEQ ID NO: 17) FYTXIPHXFGX3PP, (pattern 4; SEQ ID NO: 18) AND KX3LX: LXDIEXAX2L (pattern 5; SEQ ID NO: 19), in which (D / E) describes the alternating occupation of this sequential position by D or E, (I / L) describes the alternating occupation of this sequential position by I or L, b is the integer 10 or 11, and the X radicals are, independently of each other, any amino acid. It is ideally preferred that these patterns 1 to 5 are all present in the given sequence, with pattern 1 being closest to the N-terminus. The aforementioned PARP signature pattern is followed in the proteins according to the invention by at least one of the following standards: GXvLXEVALG (pattern 6; SEQ ID NO: 20) GX.SX, GX: .PX, LXGX: V (pattern 7; SEQ ID NO: 21) and E (Y / F) X: YXJQX.! YLL (pattern 8; SEQ ID NO: 22), in which (Y / F) describes the alternating occupation of this sequential position by Y or F, a is equal to 7 or 9 and X is in each case any amino acid. It is ideally preferred that the three C-terminal patterns are all present and in the given sequence, with pattern 8 being closest to the end C-terminal. A preferred PARP structure according to the invention can be described schematically as follows: Patterns 1 to 5 / signature of PARP / patterns 6 to 8 or Patterns 1 to 5 / zipper of leucine / signature of PARP / patterns 6 to 8, It being possible that other amino acid residues, such as, for example, up to 40, are disposed between the individual patterns and that other amino acid residues, such as, d i *? »Láb? ~ ?? * for example, up to 80, are available at the N-terminal end and / or at the C-terminal end. The PARP homologs that are particularly preferred according to the invention are human PARP2, human PARP3, mouse PARP3 proteins and functional equivalents thereof. The protein called human PARP2 contains 570 amino acids (compare with SEQ ID NO: 2). The protein called human PARP3 probably exists in 2 forms. Type 1 contains 533 amino acids (SEQ ID NO: 4) and type 2 contains 540 amino acids (SEQ ID NO: 6). The forms can arise through a different initiation of raslación. The protein called mouse PARP3 exists in 2 forms that differ from each other by the suppression of 5 amino acids (15 bp). Type 1 contains 533 amino acids (SEQ ID NO: 8) and type 2 contains 528 amino acids (SEQ ID NO: 10). The PARP homologs of the present invention differ significantly in their sequences with respect to said PARP protein of Arabidopsis thaliana (see above). For example, PARP2 and PARP3 do not contain the specific peptide sequence of PARP plant AAVLDQ IPD, corresponding to amino acid residues 143 to 152 of the Arabidopsis protein. The invention also relates to the ligands of the PARP homologs according to the invention. These ligands are preferably selected from: - * "---" - a) Antibodies and fragments of Iso itself such as, for example, Fv, Fav, F (ab ') 2 b. protein-like compounds that interact, for example, through the region of the leucine zipper or other sequential section, with the PARP, and c. "low molecular weight effectors that modulate a biological function of PARP such as, for example, the catalytic activity of PARP, ie, ADP ribosylation with NAD consumption", or binding to an activating protein or DNA. The invention also relates to nucleic acids comprising: a. a nucleotide sequence coding for at least one homologue of PARP according to the invention, or its complementary nucleotide sequence; b. a nucleotide sequence that hybridizes with a sequence as specified in a., preferably under restricted conditions; or c. nucleotide sequences that derive from the nucleotide sequences defined in a. and b. through the degeneration of the genetic code. Suitable nucleic acids according to the invention comprise in particular at least one of the partial sequences coding for the sequence patterns of amino acids mentioned.

Preferred nucleic acids according to the invention comprise nucleotide sequences such as those shown in SEQ ID N0: 1 and 3, and, in particular, partial sequences thereof which are characteristic of the PARP homologs according to the invention, such as, for example, nucleotide sequences comprising: a. nucleotides +3 to +1715 shown in SEQ ID N0: 1; b. nucleotides +242 to +1843 which are shown in SEQ ID NO: 3; c. nucleotides +221 to +1843 shown in SEQ ID NO: 5; d. nucleotides +112 to +1710 which are shown in SEQ ID NO:; or e. nucleotides +1 to +1584 shown in SEQ ID NO: 9 or partial sequences of a., b., c, d. and e. They encode the amino acid sequence patterns characteristic of the PARP homologs according to the invention. The invention also relates to expression cassettes comprising at least one of the nucleotide sequences described above according to the invention under genetic control of regulatory nucleotide sequences. These can be used to prepare recombinant vectors according to the invention, such as, for example, viral vectors of plasmids, comprising at least one expression cassette according to the invention. I The recombinant microorganisms according to the invention are transformed with at least one of the aforementioned vectors.

The invention also relates to transgenic mammals transfected with a vector according to the invention. * The invention also relates to a method for in vitro detection, for inhibitors of PARP, which can be carried out homogeneously or heterogeneously, comprising: a. incubating a poly-ADP-ribosylated blank unsupported or supported with a reaction mixture comprising: a) a PARP homologue according to the invention; a2) a PARP activator; and 10 a3) a PARP inhibitor or a substance under analysis in which at least one PARP inhibitor is suspected; b. performing the poly-ADP-ribosylation reaction; and c. qualitative or quantitative determination of the poly-ADP ribosylation of the blank. The detection method is preferably carried out by preincubating the homologue of PARP with the active PARP and the PARP inhibitor or a substance under analysis in which at least one PARP inhibitor is suspected, for example for 1-30 minutes, before carrying out the poly-ADP reaction ribosylation. After activation by DNA with single-strand breaks (termed 'activated DNA' according to the invention), the PARP poly-ADP ribosylates a large number of nuclear proteins in the presence of NAD. they include, on the one hand, the PARP itself, but also histones, etc. The poly-ADP-ribosylated target preferably used in the detection method is a histone protein in its native form or an equivalent poly-ADP-ribostable derivative derived therefrom. By way of example a histone preparation provided by Sigma (SIGMA, catalog No. H-7755; histone type II of calf thymus, Luck, JM, et al., Biol. Chem., 233, 1407 (1958) was employed. ), Satake K., et al., J. Biol. Chem. 235, 2801 (1960)). It is possible in principle to use all kinds of proteins or parts of them capable of undergoing poly-ADP-ribosylation by PARP. These are preferably nuclear proteins, such as for example histones, DNA polymerase, telomerase or PARP itself. Synthetic peptides derived from the corresponding proteins can also act as targets. In the ELISA test according to the invention it is possible to use ratios of histones in the range ranging from about 0.1 μg / well to about 100 μg / well, preferably between about 1 μg / well and about 10 μg / well. The PARP enzyme proportions range from about 0.2 pmol / cavity to about 2 nmol / cavity, preferably between about 2 pmol / cavity and about 200 pmol / cavity, the reaction mixture in each case comprising 100 μg. /cavity. Reductions to cavities of smaller sizes and, correspondingly, lower reaction volumes are possible. In the HTRF assay according to the invention, identical proportions of PARP are used, and the ratio of histone or modified histones is in the range ranging from approximately 2 ng / cavity and approximately 25 μg / cavity, preferably between approximately 25 ng / cavity and about 2.5 μg / well, the mixture comprising reaction in each case 50 μl / well. Reductions to cavities of smaller sizes and, correspondingly, lower reaction volumes are possible. The PARP activator used according to the invention is preferably activated DNA. Several types of damaged DNA can function as activators. Damage to DNA can be caused by digestion with Dnasas or other DNA-modifying enzymes (eg, restriction endonucleases), by irrationality or other physical methods or DNA guiding treatment. It is also possible to simulate the DNA damage situation of. a targeted manner, employing synthetic oligonucleotides. In the tests indicated by way of example, activated calf thymus DNA was used (Sigma, product No. D4522; CAS: 91080-16-9, prepared by method 25). ¡__¿__ _ | __i_S ______ _____ | _fflÍ ^ r-yji t -, your Jf «a * í t t A '* ij4i} feafr1 from Aposhian and Kornberg using calf thymus DNA (SIGMA D-1501) and deoxyribonuclease type I (D-4263). Aposhian H. V. and Kornberg A., J. Biol. Chem., 237, 519 (1962)). The activated DNA was used in a concentration range between 0.1 and 1000 μg / ml, preferably between 1 and 100 μg / ml, in the reaction step. The poly-ADP-ribosylation reaction is initiated in the method according to the invention by the addition of NAD +. The NAD concentrations were in the range of about 0.1 μM to about 10 inM, preferably in the range of about 10 μM to about 1 mM. In the variant of the aforementioned method which can be carried out in heterogeneous form, the poly-ADP-ribosylation of the supported target is determined using antipole antibodies (ADP-ribose). To do this, the reaction mixture is separated from the supported blank, washed and incubated with the antibody. This antibody itself may be labeled. However, as an alternative to detect an anti-poly (ADP-ribose bound) antibody, a labeled secondary secondary antibody or a corresponding labeled antibody fragment can be applied. Appropriate labels are, for example, labeled radio, labeled with chromophores or fluorophores, biotinylation, chemiluminescent labeling, . ¿Marking with a material for magnetic or, in particular, marked with enzymes, for example with horseradish peroxidase * spicy. Suitable detection techniques are generally known to those skilled in the art. In the variant of the mentioned method that can be carried out homogeneously, the unsupported blank is marked with an acceptor fluorophore. The target used preferentially in this case is the biotinylated histone, the acceptor fluorophore being coupled via avidin or streptoavidin to the biotin groups of histone. In particular, phycobiliproteins (for example phycocyanins, phycoerythrins), such as ficosianin-R (R-PC), allophycocyanin (APC), phycoerythrin-R (R-PE), phycocyanin-C (C), are suitable as acceptor fluorophores. -PC), phycoerythrin-B (B-PE) or their combinations with each other or with fluorescent dyes such as Cy5, Cy7 or Texas Red (Tandem system) (Thammapalerd, N. et al., Southeast Asian Journal of Tropical Medicine & Public Health, 27 (2): 297-303 (1996), Kronick, MN et al Climcal Chemistry, 29 (9), 1582-1586 ^ (1986); Hicks, JM, Human Pathology, 15 (2), _ 112-116 (1984)). The XL665 dye used in the examples is a cross-linked allophycocyanin (Glazer, AN, Rev. Microbil., 36, 173-198 (1982); Kronick, MN, J. Imm. Meth., 92, 1-13 (1986); MacColl, R. et al., Phycobiliproteins, CRC Press, Inc., Boca Raton, Florida (1987); MacColl, R. et al., Arch. Biochem. Biophys., 208 (1), 42-48 (1981)). It is further preferred in the homegenic method to determine the poly-ADP-ribosylation of the unsupported blank using an anti-poly (ADP-ribose) antibody that is labeled with a donor fluorophore that can transfer energy to the acceptor fluorophore when the donor and acceptor they are close in space due to the binding of the labeled antibody to the poly-ADP-ribosylated histone. A europium cryptate is preferably used as the donor fluorophore for the anti-poly (ADP-ribose) antibody. Apart from the europium cryptate used, other compounds are also possible as potential donor molecules. This may involve, on the one hand, modification of the cryptate cage. The replacement of europium by other metals from the group of rare earths such as terbium is also feasible. It is crucial that the fluorescence has a long duration to guarantee the temporary delay (López, E. et al., Clin.Chem./2/19/1920-2013).; U.S. Patent 5,534,622). The detection methods described above are based on the principle that there is a correlation between the activity of I PARP and the amount of ADP-ribose polymers formed on the histones. The assay described here makes it possible to quantify the ADP-ribose polymers using specific antibodies in the form of an EL ESA assay and an HTRF assay (homogeneous fluorescence resolved over time). Specific embodiments of these two assays are described in detail in the following examples. The test method developed HTRF (time-resolved homogeneous fluorescence) measures the formation of poly (ADP-ribose) on histones using specific antibodies. In contrast to the ELISA, this test is carried out in homogeneous phase without separation or washing steps. This enables a greater sample throughput and less susceptibility to errors. The HTRF is based on the fluorescence resonance energy transfer (FRET) between two fluorophores. In a FRET assay, an excited donor fluorophore can transfer its energy to an acceptor fluorophore when both are close in space. In the HTRF technique, the donor fluorophore is a europium [(Eu) K] cryptate and the acceptor is XL665, a stabilized allophycocyanin. The europium cryptate is based on studies carried out by Jean Marie Lehn (Strasbourg) (López E. et al., Clin. Chem. 39/2, _196-201 (1993), US Patent 5,534,622). In a homogeneous test, all the components are also present during the measurement. While this has advantages to perform the test (speed, complexity), it is necessary to avoid interference from components of the test. »^^^^^^^^^^ _ ^^^^^^ * ^^^^^^ _ ^ _ ^^^ * ^^^^^^^^^ * ^^^^^^^^ ^^^^^^^^^ ||||| 1 ^^^^ _ ^^^ S (^^^ _ ^ ^^^^^^^^^ ess (inherent fluorescence, extinction by dyes, etc. The HTRF avoids this interference by delayed measurement at two wavelengths (665 nm, 620 nm) .The HTRF has a very long decay time and therefore a delayed measurement is possible. from short-lived background fluorescence (for example, from test components or inhibitors of the substance library) .In addition, the measurement is always carried out at two wavelengths in order to compensate for the effects of extinction of the substances The HTRF assays can be carried out, for example, in a 96- or 384-well microtitre plate format and evaluated using a microplate analyzer for HTRF (Canberra Packard) discovery.They are also provided in accordance with the invention following met Methods of in vitro identification for PARP ligands, particularly for a PARP homologue according to the invention. A first variant is effected by: al) immobilization of at least one homologue of PARP on a support; bl) contacting the immobilized PARP homologue with a substance under analysis in which at least the presence of a ligand is suspected; and cl) determination, when appropriate after an incubation period, of the constituents of the substance under analysis bound to the homologue of immobilized PARP. A second variant comprises: a2) immobilization on a support of a substance under analysis comprising at least one possible ligand for the homologue of PARP; b2) contacting the substance under immobilized analysis with at least one PARP homologue for which a ligand is sought; and c2) examination of the substance under immobilized analysis, where applicable after an incubation period, for the union of the PARP homologue. The invention also relates to a method for the qualitative or quantitative determination of a nucleic acid encoding a PARP homologue, comprising: a) incubation of a biological sample with a defined amount of an exogenous nucleic acid according to the invention (for example with a length of about 20 to 500 bases or greater), hybridization, preferably under restricted conditions, determination of the nucleic acids which are hybridized and, when appropriate, comparison with a standard; or b) incubation of a biological sample with a defined quantity of pairs of oligonucleotide primers with specificity for a nucleic acid encoding a PARP homologue, amplification of the nucleic acid, determination of the amplified product and, where appropriate, comparison with a standard. The invention also relates to a method for the qualitative or quantitative determination of a homologue of PARP according to the invention, comprising: a) incubation of a biological sample with at least one associated molecule specific for a homologue of PARP, b) detection of the ligand / PARP complex and, where appropriate, c) comparison of the result with a standard. The ligand in this case is preferably an anti-PARP antibody or a combinable fragment thereof, which carries a detectable label when appropriate. The methods of determination according to the invention for PARP, particularly for PARP homologs and for the nucleic acid sequences modifying them, are suitable and advantageous for the diagnosis of tissue damage related to sepsis or ischemia, particularly strokes, myocardial infarctions, diabetes or septic shock. The invention also comprises a method, for determining the effectiveness of the PARP effectors, comprising: a) incubation of a PARP homologue according to the invention with a substance ba or analysis comprising an effector of physiological or pathological PARP activity; removal of the effector again when appropriate; and b) determination of the activity of the PARP homologue, when appropriate, after adding substrates or co-substrates. The invention also relates to gene therapy compositions which contain in a gene-acceptable carrier vehicle a nucleic acid construct comprising: an antiparallel nucleic acid versus a coding nucleic acid according to the invention; or b) a ribozyme versus a non-coding nucleic acid according to the invention; or c) codes for a specific PARP inhibitor. The invention also relates to pharmaceutical compositions comprising, in a pharmaceutically acceptable carrier, at least one PARP protein according to the invention, at least one molecule associated with PARP according to the invention or at least one coding nucleotide sequence according to the invention. the invention. Finally, the invention relates to the use of ligands of a PARP homologue for the diagnosis or therapy of pathological conditions in whose development and / or advancement is involved at least one PARP protein, in particular a PARP homologue according to the invention, or a polypeptide derived therefrom. The ligand used can be, for example, a associated molecule of low molecular weight whose molecular weight can be, for example, less than about 2000 dalton or less than about 1000 dalton. The invention further relates to the use of PARP ligands for the diagnosis or therapy of pathological conditions mediated by an energy deficit. An energy deficit is, for the purposes of the present invention, in particular, a cellular energy deficit that will be observed in the patient systemically or in individual regions of the organism, organs or parts of organs, or tissues or parts of tissues. This is characterized by a decrease in NAD and / or ATP that goes beyond (above or below) the physiological range of variation of the level of NAD and / or ATP, and is mediated preferentially by a protein with PARP activity, particularly a PARP homologue according to the invention, or a polypeptide derived therefrom. The 'energy-deficit-mediated disorders', for the purposes of the invention, additionally comprise those in which the tissue damage is attributable to cell death resulting from necrosis or apoptosis The methods according to the invention are suitable for the treatment and prevention of the resulting tissue damage, of cellular damage due to apoptosis or necrosis, damage to nervous tissue due to ischemia and / or reperfusion, neurological disorders, neurodegenerative disorders, vascular effusion, for the treatment and prevention of cardo-vascular disorders, for the treatment of other disorders or conditions such as, for example, macular degeneration due to aging, AIDS or other immunodeficient disorders, arthritis, atherosclerosis, cachexia, cancer, degenerative disorders of the skeletal muscles, diabetes, cranial trauma, inflammatory disorders of the gastrointestinal tract such as, example, Crohn's disease; muscle; osteoarthritis; osteoporosis; chronic and / or acute pain; renal insufficiency; retinal ischemia; septic shock (such as, for example, endotoxic shock); aging of the skin or aging in general; general manifestations of aging; The methods according to the invention can additionally be used to prolong the life and the proliferative capacity of the body cells and to sensitize tumor cells in relation to irradiation therapies. The invention relates in particular to the use of a molecule associated with PARP as previously defined for the diagnosis or therapy (acute or prophylactic) of pathological states mediated by energy deficits and selected from: neurodegenerative disorders, or tissue damage caused by sepsis or isgemia , "particularly from .., k. ij 1 neurotoxic disturbances, strokes, myocardial infarction, damage during or after lysis of infarction (for example with TPA, Reteplace or mechanically with laser or Rotablator) and micro-infarcts during and after a heart valve replacement, resections of aneurysms and cardiac transplants, traumas in head and spine, renal infarctions (acute renal failure, acute renal failure or damage during and after renal transplantation), skeletal muscle damage, hepatic infarcts (liver failure, damage during or after a liver transplant), peripheral neuropathies, AIDS dementia, septic shock, diabetes, neurodegenerative disorders after ischemia, trauma (craniocerebral trauma), massive hemorrhage, subarachnoid hemorrhages and stroke, as well as neurodegenerative disorders such as Alzheimer's disease, dementia by multi-infarct, Huntington's disease, Parkinson's disease, lateral sclerosis I amiotrópica, epilepsy, especially of attacks of generalized epilepsy such as petit mal and tonoclinic attacks and partial epileptic seizures, such as the temporal lobe, and complex partial seizures, renal failure, also in the chemotherapy of tumors and prevention of metastasis and for the treatment of inflammations and rheumatic disorders, eg. of rheumatoid arthritis; also for the treatment of revascularization of coronary arteries with critical narrowing and peripheral arteries with critical narrowing, eg. arteries of the legs. 'Ischemia', for the purposes of the invention, comprises a localized subprovision of oxygen to a tissue, caused by blockage of arterial blood flow.The global ischemia occurs when blood flow to the entire brain is interrupted for a limited period of time. This can be caused, for example, by cardiac arrest, focal ischemia occurs when part of the brain is interrupted by normal blood supply, focalized ischaemia can be caused by thromboembolic closure of a blood vessel, by brain trauma, edema or brain tumor Even transient ischemias can lead to far-reaching neuronal damage, although damage to nerve tissue can occur days or weeks after the onset of ischemia, some permanent damage occurs (eg, necrotic cell death). the first few minutes after the interruption of the blood supply. This damage is caused, for example, by the neurotoxicity of glutamate and follows a secondary reperfusion, such as, for example, free radical release (eg free oxygen radicals, free radicals of NO). Ischemias can also occur in other organs and tissues such as, for example, in the heart (myocardial infarction and other cardiovascular disorders caused by occlusion of the coronary arteries) or in the eye (ischemia of the retina). The invention additionally relates to the use of an effective therapeutic amount of a PARP ligand to influence neuronal activity. The 'neuronal activity', for the purposes of the invention, may consist in the stimulation of damaged neurons, promotion of neuronal regeneration or treatment of neuronal disorders, 'neuronal damage', for the purposes of the invention, comprises all kinds of damage to the "nervous tissue" and any physical or mental deterioration or death resulting from this damage The cause of the damage may be, for example, of a metabolic, toxic, chemical or thermal nature and includes as an example ischemia, hypoxia, trauma, cerebrovascular damage, operations, pressure, haemorrhages, irradiation, vasospasm, neurodegenerative disorders, infections, epilepsy, disorders of perception, alterations of glutamate metabolism and the side effects caused by all this. 'Nervous tissue', for the purposes of the invention , comprises the various components that make up the nervous system, formed, among others, by neurons, glia cells, astrocytes, Schwann cells, the system Ascular that 'is found inside and for supply, the Central Nervous System, brain, brain stem, spine, peripheral nervous system, etc.

'Neuroprotector', for the purposes of the invention, comprises the reduction, cessation, retardation or increase of neuronal damage and the protection, restoration and regeneration of nervous tissue that was exposed to neuronal damage. 'Prevention of neurodegenerative disorders' includes the possibility of prevent, delay and increase neurodegenerative disorders in people who have been diagnosed or who are included in groups at risk for these degenerative disorders. Treatments for people who already suffer symptoms of these disorders are included in the same way. "Treatment", for the purposes of the invention, comprises: (i) prevention of a disorder, disturbance or condition in persons predisposed to them; (ii) prevention of a disorder, disturbance or condition due to delay of its progress, and (iii) increase of a disorder, a disturbance or a condition Examples of 'neurological disorders' that can be treated by the methods according to the invention are: neuralgia (trigeminal, glossopharyngeal), iastenia gravis, muscular dystrophies, lateral to iophic sclerosis (ALS), progressive muscular atrophy, peripheral neuropathies caused by poisoning (eg lead poisoning), Guillain-Barré syndrome, Huntington's disease, Alzheimer's disease, Parkinson's disease, or disorders of the plexus. The. Methods according to the invention are preferably suitable for treating neurological disorders selected from: cranial trauma such as, for example, a traumatic brain injury; physical damage to the spine; stroke associated with brain damage, such as vascular effusion in combination with hypoxia and brain damage, and damage "by cerebral reperfusion, disorders caused by demyelination myelopathies, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis). Methods according to the invention can additionally be used to treat cardiovascular disorders. 'Cardiovascular disorders', for the purposes of the invention, include those which cause ischemia or which are caused by ischemia or by ischemia / reperfusion of the heart. the coronary arteries (for example atherosclerosis), angina pectoris, myocardial infarction, cardiovascular damage due to cardiac arrest or bypass operation. The methods according to the invention can be used for the treatment of cancer or for sensitizing cancer cells for radical therapy. The term "cancer" will be understood in its broadest sense The protein modulators according to the invention can be used as 'anti-cancer therapeutic agents'. For example, the methods can be used to treat cancers or tumor cells, such as ACTH-generating tumors, acute lymphatic or lymphoblastic leukemia; acute or chronic lymphocytic leukemia; acute non-lymphocytic leukemia; bladder cancer; brain tumors; breast cancer; cervical carcinoma; chronic myelocytic leukemia; bowel cancer; T-zone lymphoma; endometriosis; esophagus cancer; gallbladder cancer; Ewing's sarcoma; Head and neck cancer; Tongue cancer; Hodgkin's lymphoma; Kaposi's sarcoma; Kidney cancer; Liver cancer; lung cancer; mesothelioma; multiple myeloma; neuroblastoma; Non-Hodgkin's lymphoma; osteosarcoma; ovarian carcinoma; glioblastoma; mammary carcinoma; cervical carcinoma; prostate cancer; pancreatic cancer; penis cancer; retinoblastoma; skin cancer; stomach cancer; thyroid cancer; uterine carcinoma; vaginal carcinoma; ilm tumor; or trophoblastoma. "Radiosensitizer" or "radiation sensitizer", for the purposes of the invention, refers to molecules that increase the sensitivity of the cells of the body to irradiation with electromagnetic rationing (for example X-rays) or accelerate this treatment by irradiation. Radiation sensitizers increase the sensitivity of cancer cells to the toxic effects of electromagnetic radiation. Those described in the literature include mitomycin C, 5-bromo-deoxyuridine and metronidazole. Is ___ ^^^^^ ____ ^^^^ _ ^^^^^ and fe ^^^ £ ^^ ______ ^ __ ^^^^ _ ^ _ ^^^^ a__s »^ l» | ___ Is it possible to use radiation with wavelengths in the range of 10"" to 10 meters, preferably gamma rays (10 ~ 20 to 10"13 m), X-rays (10" to 10"5 m), ultraviolet radiation (10 nm to 400 nm) , visible light (400 nm to 700 nm), infrared radiation (700 nm to 1 mm) and microwave radiation (1 mm to 30 cm). the disorders that can be treated by such therapy are, in particular, neoplastic disorders, benign tumors . or malignant and cancer treatment of other disorders using electromagnetic radiation is also possible the invention will now be described in greater detail with reference to the accompanying figures These illustrate:.. in Figure 1 a sequential alignment of human PARP (human PARP1) and two preferred PARPs according to the invention (human PARP2, human PARP3, murine PARP3), sequential matches between human PARP1 and human PARP2, human PARP3 or PA RP3 of murine stand out boxed. The majority sequence is indicated above the alignment. The zinc finger patterns of human PARP1 are located in the sequential sections corresponding to amino acid residues 21 to 56 and 125 to 162; In Figure 2 Northern diagrams with various human tissues to illustrate the tissue distribution of the PARP2 and PARP3 molecules according to the invention. Route 1: brain; travel 2: heart; travel 3: skeletal muscle; travel 4: color; travel 5: time- travel 6: spleen; travel 7: kidney; travel 8: liver; travel 9: intestine; route 10: placenta: route 11; lung; 'route 12: peripheral blood leukocytes; the respective position of the size standard (kb) is indicated. In Figure 3 a Northern diagram with several other human tissues to illustrate the tissue distribution of the PARP3 molecule according to the invention. Route: 1: heart; travel 2: brain; travel 3: placenta; travel 4: lung; travel 5: liver; travel 6: skeletal muscle; travel 7: kidney; travel 8: pancreas; the respective position of the size standard (kb) is indicated. In Figure 4 a Western diagram with various human tissues to illustrate the tissue distribution of the molecule PARP3 according to the invention at the protein level. Route 1: heart; travel 2: lung; travel 3: liver; travel 4: spleen; travel 5: kidney; travel 6: colon; travel 7: muscle; travel 8: brain; the respective position of the size standard (kb) is indicated. In Figure 5 a Western diagram with various human tissues to illustrate the tissue distribution of the PARP3 molecule according to the invention. Route 1: frontal cortex; travel 2: posterior cortex; travel 3: cerebellum; travel 4: hippocampus; travel 5: bulb olfactory; travel 6: striatum; travel 7: thalamus; j ^ M ^^^ path .8: middle brain; travel 9: endonasal cortex; travel 10: bridge; travel 11: marrow; travel 12: spine. In Figure 6 a diagrammatic representation of the PARP (ELISA) assay. In the Fifura 7 a diagrammatic representation of the PARP assay (HTRF). Other preferred embodiments of the invention are described in the following sections. PARP homologues and functional equivalents Unless otherwise indicated, for the purposes of the present disclosure, amino acid sequences are indicated beginning with the N-terminus. If the one-letter code for amino acids is used, then G is glycine, A is alanine, V is valine, L is leucine, I is isoleucine, S is serine, T is threonine, D is aspartic acid, N is asparagine, E is glutamic acid, Q is glutamine, W is tryptophan, H is histidine, R is arginine, P is proline, K is lysine, Y is tyrosine, F is phenylalanine, C is cysteine and M is methionine. The present invention is not confined to the PARP homologs specifically described above. Conversely, those homologs that are functional equivalents thereof are also included. Functional equivalents comprise variants of the proteins specifically described herein, both natural, such as, for example, species-specific or organ-specific, as well as those produced artificially. Functional equivalents according to the invention differ by addition, substitution, inversion, insertion and / or deletion of one or more amino acid residues of human PARP2 (SEQ ID NO: 2), human PARP3 (SEQ ID NO: 4 and 6) and mouse PARP3 (SEQ ID NO: 8 and 10), there being at least retention of the NAD binding function of the protein mediated by a C-terminal catalytic functional domain. Similarly, the catalytic activity of poly (ADP-ribose) production should preferably be retained. Functional equivalents also comprise, where appropriate, those variants in which the region similar to the leucine zipper is essentially retained. It is also possible, for example, starting with the sequence for human PARP2 or human PARP3, to replace certain amino acids with those with similar physicochemical properties (size, basicity, hydrophobicity, etc.). It is possible, for example, that arginine residues be replaced by lysine residues, valine residues by isoleucine residues or aspartic acid residues by glutamic acid residues. However, it is also possible that one or more amino acids are exchanged together, added or deleted, or several of these operations are ^^^^ j ^^^^^ S * ^ ^ _ ^^ _ s ^ e ^^^^^^^^ S ^^^^^ g ^^^^^^^ * S ^ * y ^ ^^^^ i? ^ «j-lfajt u? AA combine together. Proteins that have been modified in this way from the sequence of human PARP2 or human PARP3 have at least 60%, or better, at least 75%, or, better yet, at least 85% homology with the sequence of departure, calculated using the algorithm of Pearson and Lipman, Proc. Natl. Acad. Sci ("USA) 85 (8), 1988, 2444-2448 The following amino acid level and DNA level homologies have been determined between human PARP1, 2 and 3 (FastA, Pearson and Lipman, loe. Cit. Program). : Homologies of amino acids: The numbers in parentheses indicate the number of overlapping amino acids. DNA homologies: The numbers in parentheses indicate the number of overlapping nicleotides.

The polypeptides according to the invention can be classified as homologous poly (ADP-ribose) polymerases on the basis of great similarity in the region of the catalytic domain. It is also essential for the invention that novel PARP homologs have no conventional zinc finger patterns. This means that these enzymes are not necessarily involved in DNA repair or are in a way that differs from PARP1, but they are still able to develop their pathological mechanism (NAD consumption and therefore energy consumption due to ATP consumption). ). The strong protein expression, particularly of PARP3, observable in the Western diagram suggests a significant role in NAD consumption. This is particularly important for the development of drugs. The novel potential inhibitors of the polymerases according to the invention can thus inhibit the pathological functions without having adverse effects on the desired physiological properties. This was impossible with the inhibitors of PARPs known until now as there was always also inhibition of DNA repair function. The potentially mutagenic effect of the known PARP inhibitors is thus easy to understand. It is also conceivable to design PARP inhibitors so that they efficiently inhibit all homologs i ^ í ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ ^ '^^^ i ^^^ t ~ k + n? ? * * J &Bf &JLf of PARP with high affinity. In this case, an enhanced effect is conceivable where appropriate. The homologue of PARP that is preferred according to the invention and shown in SEQ ID NO: 2 (human PARP2) can be advantageously isolated from human brain, heart, skeletal muscle, kidney and liver. The expression of PARP2 in other tissues or organs is distinguished by being weaker. The homologue of PARP which is preferred according to the invention and shown in SEQ ID NOM and 6 (human PARP3) can be advantageously isolated from the brain (in this case very preferably from the hippocampus), heart, skeletal muscle, liver or kidney. humans. The expression of PARP3 in other tissues or organs, such as muscle or liver, is distinguished by being weaker. The expert familiar with protein isolation will make use of the combination of preparative methodologies that is most suitable in each case to isolate natural PARPs according to the invention from tissues, or PARPs prepared by recombination according to the invention from cell cultures. 'Standard preparative methods are described, for example, in Cooper, T.G., Biochemische Arbeitsmetoden, published by Walter de Gruyter, Berlin, New York or in Scopes, R. Protein Purification, Springer Verlag, New York, Heidelberg, Berlin.

The invention further relates to homologs of PARP2 and PARP3 which, although they may be isolated from other eukaryotic species, ie invertebrates and vertebrates, I especially other mammals such as, for example, mice, rats, cats, dogs, pigs, sheep, cows, horses or monkeys, or other organs such as, for example, the myocardium, have the essential structural and functional properties predetermined by the PARPs according to the invention. In particular, human PARP2 that can be isolated from human brain, and their functional equivalents, are preferred agents to develop inhibitors of neurodegenerative diseases such as stroke. This is because it can be assumed that the development of drugs based on PARP2 as an indicator makes it possible to develop inhibitors optimized for use in the human brain. However, it can not be ruled out that the inhibitors developed on the basis of PARP2 can also be used for the treatment of pathological conditions mediated by PARP also in other organs (see tissue distribution of the proteins according to the invention). PARP2 and presumably PARP3, similar to what occurs with PARP1, are activated by damaged DNA, although presumably by a different mechanism. Its importance in DNA repair is imaginable. The blocking of PARPs ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ "with the invention would also be beneficial in situations such as cancer (eg in the radiosensitization of patients with tumors.) Another essential biological property of the PARPs according to the invention and their functional eguivalents will be in their ability to join an interactive partner molecule The human PARP2 and 3 differ from the previously described PARPs from higher eukaryotes such as, in particular, mammals, by having potential leucine zipper patterns.This is a typical pattern for protein-protein interactions. It is possible that these patterns allow the modulation of PARP activity by an interactive associated molecule.Thus, this additional structural element provides a possible starting point for the development of PARP effectors such as, for example, inhibitors. it also refers to proteins that interact with PARP2 and / or 3, preferably those that produce their activation or inactivation. The invention also relates to proteins that still have the aforementioned ligand-binding activity and that can be prepared starting from the amino acid sequences specifically described, by directed modifications.

It is possible, starting from the peptide sequence of the proteins according to the invention, to generate synthetic peptides that are used, alone or in combination, as antigens to produce polyclonal or monoclonal antibodies. It is also possible to use the PARP protein or fragments thereof to generate antibodies. Thus, the invention also relates to peptide fragments of PARP proteins according to the invention which comprise characteristic partial sequences, in particular those oligo- or polypeptides comprising at least one of the mentioned sequential standards. Fragments of this type can be obtained, for example, by proteolytic digestion of PARP proteins or by chemical synthesis of peptides. Specific novel ligands for PARP2 and PARP3 Active inhibitors and preferably selective proteins were developed according to the invention, using the evaluation systems described above for ligands of PARP2 and PARP3. These inhibitors are optionally also active against PARP1. The inhibitors provided according to the invention have a strong inhibitory activity of PARP2. The values of Kx can in this case be less than approx. 1000 nM, such as minors of approx. 700 nM, less than approx. 200 nM or .ta? T .m - Í.á,? T .. -jí -ÁÉrí, j ... u? Üu. ^? .... i -. «..i-t, __ > »». «IÍHtti.-J« «t.-WfcJ ^ U.J * ^ A¿ < _il > minors of -aprox. 30 nM, for example between approximately 1 and 20 nM. The inhibitors according to the invention can also have a surprising selectivity for PARP2. This is revealed by the ratio Ki (PARPl): Ki (PARP2) for said inhibitors according to the invention, which is, for example, greater than 3 or greater than 5, such as greater than 10 or greater of 20. An example to be mentioned is 4- (N- (4-hydroxy-phenyl) aminomethyl) - (2H) -dihydrophthalazin-1-one. The preparation of this and other analogous compounds can be carried out according to Puodzhyunas et al., Pharm. Chem. J. 1973, 7, 566 or Mazkanowa et al., Zh. Obshch. Khim., 1958, 28, 2798, or Mohamed et al., Independently. J. Chem. B., 1994, 33, 769, all of which are incorporated by reference. The above-identified compound exhibits a Ki value of 113 nM for PARP2 and is eight times more selective for PARP2 than for PARP3. Nucleic acids encoding PARP homologs: Unless explicitly stated otherwise, the nucleotide sequences are indicated in the present invention in the 5 'to 3' direction. The invention also relates to nucleic acid sequences encoding the above mentioned proteins, in particular those having the given amino acid sequence ^ MB ^ Mk for SEQ ID NO: 2, 4, 6, 8 and 10, but without being restricted to them. The nucleic acid sequences that can be used according to the invention also comprise allele variants which, as described above for the amino acid sequences, can be obtained by deletion, inversion, insertion, addition and / or substitution of nucleotides, preferably from the nucleotides shown in SEQ ID NO: 1, 3, 7 and 9, but with essential retention of the biological properties and biological activity of the corresponding gene obtained. The nucleotide sequences that can be used are obtained, for example, by nucleotide substitutions that cause silent changes (without alteration of the amino acid sequence) or conservative amino acid changes (exchange of amino acids of the same size, charge, polarity or solubility). The nucleic acid sequences according to the invention also encompass functional equivalents of the genes, such as eukaryotic homologs for example from invertebrates such as Caenorhabditis or Drosophila, or vertebrates, preferably from the mammals described above. Preferred genes are those from vertebrates that encode a product gene that has the essential properties of the invention described above. The nucleic acids according to the invention can be obtained in a conventional manner by various routes: For example, a genomic or cDNA library can be searched for a DNA encoding a PARP molecule or part of it. For example, a cDNA library obtained from human brain, heart or kidney can be searched with a suitable probe such as, for example, a labeled single-stranded DNA fragment corresponding to a partial sequence of suitable length selected from SEQ ID. NO: 1 or 3, or the complementary sequence of it. For this purpose, it is possible, for example, that the DNA fragments of the The library, which has been transferred to a suitable cloning vector, is plated on agar plates after being transformed into a bacterium. The clones can then be transferred to nitrocellulose filters and, after denaturation of the DNA, hybridized with the probe marked. Then the positive clones are isolated and characterized. The DNA encoding the PARP homologs according to the invention or partial fragments can also be synthesized chemically from the sequential information contained in the present application. For example, it is possible with this In order that oligonucleotides with a length of about 100 bases are synthesized and linked sequentially in a manner known per se, for example, by supporting the appropriate terminal restriction cleavage sites. The nucleotide sequences according to the invention can also be prepared with the help of the chain reaction rrr. ,.. .TO. * of the polymerase (PCR). To do this, a white DNA is hybridized, such as, for example, DNA from a suitable full-length clone, with a pair of oligonucleotide synthetic baits having a length of approx. 15 bases and that bind to opposite ends of the white DNA. The stretch of sequence that remains between them is filled with the DNA polymerase. The reiteration of this cycle often allows the amplification of the white DNA (compare with White et al (1989), Trends Genet, 5, 185). It will be understood that the nucleic acid sequences according to the invention also include truncated sequences, single-stranded DNA or RNA of the complementary sequence of coding or non-coding DNA, mRNA sequences and cDNAs derived therefrom. The invention also encompasses nucleotide sequences that hybridize to the mentioned sequences under restricted conditions. Restricted hybridization conditions, for the purposes of the present invention, exist when the hybridization sequences have a homology of between approx. 70 to 100%, "such as, for example, about 80 to 100% or 90 to 100% (preferably in a stretch of amino acids of at least about 40, such as, for example, about 50, 100, 150 , 200, 400 or 500 amino acids.) Restricted conditions for DNA search, in particular for cDNA libraries, exist, for example, when the ^ ^ ^ ^ ^ ^^^^ J ^^^^^^^^^^^^ ßfc *? ^^^^^^^ 4 ^^^^^ J ^^^ J ^^ ilfc ^^ ^^ g ^^^^^^^^^^^ »^ ^ ^ ^ ^ ^ ^ ^ Hybridization mixture is washed with 0.1X SSC buffer (20X SCC buffer = 3M NaCl, 0.3M sodium citrate, pH 7.0) and 0.1% SDS at a temperature of about 60 ° C. The Northern analysis spots are washed under restricted conditions with 0, IX SSC, 0.1% SDS at a temperature of approx. 65 ° C, for example. Nucleic Acid Derivatives and Expression Constructs: It will be understood that nucleic acid sequences also include derivatives such as, for example, alternative splice variants or promoter variants. Promoters operably linked upstream of the nucleotide sequences according to the invention can also be modified by addition (s) or substitution (s), inversion (s), insertion (s) and / or deletion (s) of nucleotides, but without altering the functionality or activity of the promoters. The promoters may see their activity increased by modification of their sequence, or be completely replaced by more efficient promoters, even from heterologous organisms. The promoter variants described above are used to prepare expression cassettes according to the invention. Specific examples of splicing variants of human PARP2 that may be mentioned are: ^^ W ^ Human PARP2a variant: Suppression of base pairs 766 to 904 (compare with SEQ ID NO: 1). This leads to a frame shift with a new cutting codon ('TAA "corresponding to nucleotides 922 to 924 in SEQ ID NO: 1) 5 Human PARP2b variant: Insertion of 5'-gta tgc cag gaa ggt cat ggg cea gca aaa ggg tet ctg - 3 'after nucleotide 204 (SEQ ID NO: 1) This prolongs the amino acid sequence by insertion: GMPGRSWASKRVS Nucleic acid derivatives also involve 1st variants whose nucleotide sequences of the region ranging from -1 to -1000 against the initial codon have been modified so that gene expression and / or protein expression is increased. In addition to the nucleotide sequence described above, the Nucleic acid constructs that can be used according to the invention comprise, in functional and operational binding, one or more other regulatory sequences, such as promoters, amplification signals, enhancers, polyadenylation sequences, origins of replication, genes informants, selectable marker genes and the like. This binding may, depending on the intended use, lead to an increase or decrease in gene expression. In addition to the novel regulatory sequences, it is possible that the natural regulatory sequence is still present in to the true structural genes. This natural regulation ^^^ M ^^^ te ^^^^ _ ^^^^^^^^ fc ^^^^^^^^^^^^^^^^^^^^^^^ ^ t ^^^ f ^^^^^ gE¿ ^^^^^^^^^^^^^^^ A5 ^^^ can, where appropriate, be extinguished by the genetic modification, and the expression of the genes increase or decrease. However, the gene construct can also have a simpler structure, ie no additional regulatory signal is inserted against the structural genes, and the natural promoter with its regulation is not deleted. In contrast, the natural regulatory sequence mutates such that regulation no longer occurs, and gene expression increases or decreases. It is also possible to insert additional advantageous regulatory elements at the 3 'end of the nucleic acid sequences. The sequences of the nucleic acids can be present in one or more copies in the gene construct. Advantageous regulatory sequences for the expression method according to the invention are present, for example, in promoters such as eos, tac, trp, tet, trp-tet, lpp, lac, lpp-lac, laclq, T7, T5, T3 , gal, tre, ara, SP6, 1-PR or the 1-PL promoter, which are advantageously used in Gram-negative bacteria. Other advantageous regulatory sequences are present, for example, in the Gram-positive promoters amy and SP02, in the yeast promoters ADC1, Mfa, AC, P-60, CYC1, GAPDH or in the plant promoters CaMV / 35S, SSU, OCS, lib4, usp, STLSl, B33, 'us in the ubiquitin or phaseolin promoter.

It is possible in principle to use all natural promoters with their regulatory sequences. It is also possible and advantageous to use synthetic promoters. Said regulatory sequences are intended to make possible the specific expression of nucleic acid sequences and protein expression. This may mean, for example, depending on the host organism, that the gene is expressed or overexpressed only after induction, or that it is expressed and / or overexpressed immediately. Regulatory sequences or factors may also preferably have a positive influence on, and thus include or diminish, expression. Thus, the increase of regulatory elements can advantageously take place at the level of transcription using strong transcription signals such as promoters and / or enhancers. However, it is also possible to increase translation, for example by improving the stability of mRNA. Enhancers means, for example, DNA sequences that produce increased expression via an improved interaction between RNA polymerase and DNA. The recombinant nucleic acid construct or gene construct, by expression in a suitable host organism, is advantageously inserted into a host-specific vector that enables optimal expression of the genes in the host. The vectors are well known to experts in -, * Z »# i ^ ¿> ^ & amp; & gj - «Aá¿¿, - * - * i * J¿k. < neither. _j ^ t: L.-._ £ i ^ ^ _ i ^ ^: ^^ ^ S ^^ * ^ l ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ : ^^^ e¡: _ ^ ^ ^ í ^ art and 'are found, for example, in' Cloning Vectors' (Pouwels PH et al., Ed., Elsevier, Amsterdam-New York-Oxford, 1985). Apart from plasmids, vectors also mean any other vector known to those skilled in the art such as, for example, phages, viruses, such as SV40, CMV, baculovirus and adenoviruses, transposons, IS elements, pseudostems, cosmids, and linear or DNA. These vectors may undergo autonomous replication in the host organism or chromosomal replication.Expression of the constructs: The recombinant constructs according to the invention described above are advantageously introduced into a suitable host system and are expressed.Familial cloning and transfection methods for those skilled in the art they are preferably used for the purpose of producing the expression of said nucleic acids in the particular system of ex Suitable systems are described, for example, in Current Protocols in Molecular Biology, F. Ausubel et al., Ed. Wiley Interscience, New York 1997. Suitable host organisms are in principle all organisms that make it possible to express nucleic acids. according to the invention, its allelic variants, its equivalents or functional derivatives or the construct? of recombinant nucleic acid. Host organisms means, for example, bacteria, fungi, yeasts, plant cells or ^? ^ ^? ¿¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡¡ Preferred organisms are bacteria such as those of the genera Escherichia, such as, for example, Escherichia coli, Streptomyces, Bacilius or Pseudomonas, eukaryotic microorganisms such as Saccharomyces cerevisiae, Aspergillus, higher eukaryotic cells from animals or plants, such as cells Sf9 or CHO. The produced gene can also, if necessary, be expressed in transgenic orgenisms such as transgenic animals, particularly in mice, sheep or transgenic plants. The transgenic organisms can also be the so-called animals and plants 'out of combat' (in the original, knock-out) in which the corresponding endogenous gene has been disconnected, such as, for example, by mutation or partial or total suppression. The combination of host organisms and appropriate vectors for organisms, such as plasmids, viruses or phages, such as, for example, plasmids with the RNA polymerase / promoter system, phages? Or other temperate phages or transposons and / or Other advantageous regulatory sequences form an expression system The phrase "expression systems" preferably means, for example, a combination of mammalian cells such as CHO cells, and vectors, such as the pcDNA3neo vector, which are suitable for mammalian cells. | ^^^^^^? ^^^^ l ^^^ t ^^? j & £ ^ & £ > As described above, the gene produced can also be advantageously expressed in transgenic animals, for example mice and sheep, or in transgenic plants. It is similarly possible to program cell-free translation systems with the RNA derived from the nucleic acid. The produced gene can also be expressed in the form of therapeutically or diagnostically suitable fragments. In order to isolate the recombinant protein it is possible and advantageous to use vector systems or oligonucleotides that prolong the cDNA by certain nucleotide sequences and thus encode modified polypeptides that serve to simplify the purification. Suitable modifications of this type are, for example, the so-called slopes (in the original, tags) that act as an anchor, such as, for example, the modification known as hexa-histidine anchor, or epitopes that can be recognized as antigens by means of of antibodies (described, for example in Harlow, E. and Lane, D., 1988, Antibodies: A Laboboratory Manual, Cold Spring Harbor (NY) Press). These anchors can be used to attach the proteins to a solid support such as, for example, a polymer matrix, which may be, for example, filling the inside of a chromatographic column, or to a microtiter plate or other support. These anchors can also be used to recognize proteins at the same time. It is also possible to use for the ___ IT? __ I __________ fc__ '----- "- recognition of conventional marker proteins such as fluorescent dyes, enzymatic labels that form a detectable reaction product after reaction with a substrate, or radioactive labels, alone or in combination with anchors to derivatize the proteins Production of antibodies: The anti-PARP2 antibodies are produced in a manner familiar to the person skilled in the art. Antibodies means polyclonal, monoclonal, human or humanized antibodies, or fragments thereof, single chain antibodies or synthetic antibodies. , in the same way as antibody fragments such as Fv, Fab and F (ab '); suitable production methods are described, for example, in Campbell, AM, Monoclonal Antibody Technology, (1987) Elseveier Verlag, Amsterdam, New York , Oxford and in Breitling, F. and Dübel, S., Rekombinante Antikórper (1997, Spektrum Akademisher Verlag, Heidelberg.) Additional use of the coding sequence: The present additional cDNA provides the basis for the cloning of the genomic sequence of the new PARP genes. It also includes the relevant regulatory sequence or promoter sequence, which is available, for example, by sequencing the 5 'upstream region of the cDNA according to the present invention or located in the introns of the genes. The sequence information ^^^^^^^^^^^ f ^^^^^^^^ X ^ £ ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ ^^^^^^^^^^^^ ^^^^^ ^ ^ DD cDNA is also the basis for producing counter-sense molecules or ribozymes, with the addition of known methods (see Jones, JT and Sallenger, BA (1997) Nat. Biotechnol., 15, 902; Nellen, W. and Lichtenstein, C . (1993) TIBS, 18, 419). Genomic DNA can also be used to produce the constructs of the genes described above. Another possibility of using part or all of the nucleotide sequence is to generate transgenic animals. Transgenic overexpression or genetic deletion of sequence information in appropriate animal models can provide additional valuable information about the (duck) physiology of the new genes. Therapeutic applications: In situations where there is a prevalent deficiency of a protein according to the invention, it is possible to employ several substitution methods. On the one hand, the protein, natural or recombinant, can be administered directly or by gene therapy in the form of its coding nucleic acid (DNA or RNA). It is possible to use any suitable vector for this, for example, viral and non-viral vehicles. Appropriate methods are described, for example, by Strauss and Barranger in Concepts in Gene Therapy (1997), Watler de Grutyer, editor. Another alternative is provided by stimulation of the endogenous gene by the appropriate agents.

It is also possible to block the production or inactivation of the PARPs according to the invention, for example, by proteases. Finally, the inhibitors or agonists of the I PARPs according to the invention can be used. 5 In situations where an excess or overactivated PARP is present, several types of inhibitors can be used. This inhibition can be achieved by counter-sense molecules, ribozymes, oligonucleotides or antibodies, and by low molecular weight compounds. The active substances according to the present invention, for example, PARP proteins, nucleic acids and ligands of PARP such as, for example, antibodies or modulators, can be administered as therapeutically active substances individually or as a mixture with other therapeutically active substances. They can be administered as such, but are generally administered in the form of pharmaceutical compositions, for example, mixtures of active substance (s) with at least one suitable pharmaceutical carrier or diluent. The active substances or compositions can be administered in any manner suitable for the particular therapeutic purpose, for example, orally or parenterally. The nature of the pharmaceutical composition and the pharmaceutical carrier or diluent depends on the required mode of administration. Oral compositions can, for example, presented in the form of tablets or capsules and may contain common excipients such as binders (eg, syrup, acacia, gelatin, sorbitol, tragacanth or polyvinylpyrrolidone), bulking agents (eg, lactose, sugar, corn starch, phosphate calcium, sorbitol or glycine), lubricants (for example, magnesium stearate, talc, polyethylene glycol or silica), disintegrants (for example, starch) or wetting agents for example sodium lauryl sulfate). The oral liquid products may be in the form of aqueous or oily suspensions, solutions, emulsions, syrups, elixirs or sprays, etc., or they may be in the form of dry powders for reconstitution with water or other suitable vehicle. The liquefied products of this type may contain conventional additives, for example, suspending agents, flavorings, diluents or emulsifiers. Suspensions or solutions with conventional pharmaceutical vehicles can be used for parenteral administration. The parenteral administration of the active substances according to the invention advantageously takes place using a liquid pharmaceutical composition which can be administered parenterally, in particular, intravenously. This preferably contains an effective amount of at least one active substance, preferably in the dissolved form, in a pharmaceutically acceptable carrier suitable for this purpose. Examples of pharmaceutical carriers suitable for these purposes are, in particular, aqueous solutions such as, for example, physiological saline, phosphate "buffer" salt, Ringer's solution, lactate Ringer's solution and the like. The composition may also contain aggregates such as antioxidants, chelating agents, or antimicrobial agents. The choice, in each case, of the dose of the active substance according to the invention, and the program of administration of the particular doses are subject to the decision of the doctor performing the treatment. This will select the appropriate dose and the program. of appropriate administration depending on the route of administration chosen, on the efficiency of the medication in each case, on the nature and severity of the disease to be treated, and on the patient's condition and response to therapy. Thus, for example, pharmacologically active substances can be administered to a mammal (human or animal) in doses of about 0.5 mg to about 10 mg per kilo of body weight per day. They can be administered in a single dose or in several doses. Non-therapeutic applications: Nucleic acids according to the invention, such as, for example, cDNA, genomic DNA, promoter, and polypeptide, and their partial fragments, can also ^^^^? ^^^^^^^^^^^^ jjj ^^ s ^^^ s ^^^^^ a ^^ i ^^^^^^ g & ^^^^^^^^^ - A ^ Ij ^^^^^^^^^ '^^ * ^^^^^ used -in the recombinant or non-recombinant for the development of several test systems. For example, it is possible to establish a test system that is suitable for measuring the activity of the promoter or the protein in the presence of a test substance. The measurement methods in this case are preferably simple, for example, colorimetric, luminometric, fluorimetric, or radioactive immunological, and preferably allow a large number of test substances measured quickly. Tests of this type are suitable and advantageous for the so-called high performance classification. These test systems allow the tested substances to be evaluated by their linkages or by their agonism, antagonism or inhibition of proteins according to the invention. Determining the amount, activity and protein distribution according to the invention, or their underlying mRNA in the human body, it can be used for diagnosis, for determination of susceptibility and for controlling certain diseases. In the same way, the cDNA sequence and the genome sequence can provide information on the genetic causes and on the predisposition to certain diseases. It is possible to use both DNA / RNA probes and antibodies of a wide variety of types for this purpose. The nucleotide sequences or their parts, according to the present invention, can also be used in the form of suitable probes for detecting point mutations, deletions or insertions. The proteins according to the invention can also be used to identify and isolate their natural ligands or interactive partners. The proteins according to the invention can also be used additionally to identify and isolate the artificial or synthetic ligands. For this purpose, the natural protein purified or prepared recombinantly can be derivatized in such a way that it presents modifications that allow its binding to support materials. The protein linkages in this form can be incubated with various analytical compounds, such as, for example, protein extracts or peptide libraries or other sources of ligands. Peptides, proteins or non-protein substances, of low molecular weight, with specific bonds, can be isolated and characterized in this way. Non-protein substance means, for example, low molecular weight chemicals that may originate, for example, from classical drug synthesis or from the so-called libraries of substances that were synthesized in combination. The protein extracts used are derived, for example, from a homogenate of plants or parts of plants, microorganisms, human or animal tissues or organs. ^^^^^^^^ É ^^^^^^^^^^^ faith ^^^^^^^ g ^^^^^^^^^^^^^^ & ^^^ ^ fc ^ * ^ * fc ^^^^^^^^^^^ ^^^^^^^^^^^^^^^ _ ^^ a ^ BFIG marmoset ligands or associated interactive methods can also be identified as yeast two-hybrid system (Fields, S. and Song, 0. (1989) Nature, 340, 245). The expression or activation banks that can be used in this case can be derived, for example, from human tissues such as, for example, brain, heart, kidney, etc. The nucleic acid sequences according to the invention and the proteins encoded by them can be used for developmental reagents, agonists and antagonists or inhibitors for the diagnosis and therapy of chronic and acute diseases associated with the expression of one of the protein sequences of according to the invention, as, for example, with its expression increased or decreased. The developed reagents, agonists, antagonists or inhibitors can, subsequently, be used to produce pharmaceutical preparations for the treatment or diagnosis of diseases. Examples of possible diseases related to this are diseases of the brain, peripheral nervous system, cardiovascular system or eyes, septic shock, rheumatoid arthritis, diabetes, acute kidney failure or cancer. The importance of the proteins according to the invention for said indications was verified using specific inhibitors in relevant animal models.

- - • - - '- ~' - - - "- - -" "" - - '-' - - - - ^^^ ^^ ^^ ^ LJ Now, the invention is illustrated in detail with reference to The following examples. Example 1: Isolation of the cDNA from PARP2 and PARP3 The present cDNA sequences were first found in a sequence analysis of cDNA clones from a cDNA library of the human brain (Human Brain 5'Strech Plus cDNA Library, # HL3002a, Clontech). Mouse PARP3 clones were isolated from a 'lambda triplex mouse brain cDNA library' (Clontech No. of order ML5004t) The sequences of those clones are described in SEQ ID N0: 1, 3, 7 and 9. Example 2: Expression of PARP2 and PARP3 in human tissues Expression of human PARP2 and human PARP3 was investigated in twelve different human tissues by Northern blot analysis. 'Northern blotches of Human Multiple Tissues (MTN ™) provided by Clontech (# 7760) -1 and # 7780-1) was hybridized for this purpose with an RNA probe.The probe was produced for transcription in vi tro of the corresponding cDNA of PARP @ human and human PARP3 in the presence of nucleotides labeled with dogoxigenin according to manufacturer's methods (BOHERINGER MANNHEIM DIG Easy Hyb order No. 1603 558, DIG Easy Hyb, method for RNA: RNA hybridization). The protocol was modified to perform the prehybridization: 2xlh adding DNA from herring sperm (10 mg / ml of solution Hybridization ion). The ^^^^^^^^^^^^^^ ^^^^^^ j ^^ j ^ ^^^^^^^^^? I ^ í ^^^^^^^^^ ^^^^^ r ^^^^^^^^^^^^^^^^^ hybridization then takes place all night, plus, herring sperm DNA (1 mg / ml of hybridization solution ). The bands were detected using the CDP-Star protocol (Boeringher Mannheim CDP-Star ™ No. Order 1685 627). After a rigorous wash, the transcript of PARP2 was detected mainly in human brain, heart, skeletal muscles, kidneys and liver. The transcript size of approximately 1.9kv corresponds to the extension of the determined cDNA (1.85 kV) (compare in Figure 2 (A)). In other tissues or organs, the expression of human PARP2 is considerably weaker. After a severe wash, the transcript of PARP3 was detected mainly in heart, brain, kidney, skeletal muscle and liver. Expression in other tissues (placenta, lung, pancreas) is notoriously weaker (compare in Figure 2 (B)). There are at least two transcripts for human PARP3, which can be presumably explained by the different polyadenylation sites or alternative linkages. Their size (approximately 2.2 kv and 2.5 kv respectively) corresponds to the extension of the determined cDNA (2.3kv). Washing was done with 0.2 x SSC / 0.2% SDS at room temperature for 2 x 15 minutes, and "then with 0.1 x SSC / 0.1% SDS at 65 ° C for 2x15 minutes (prepared 20X SSC: 3M NaCl, citrate sodium 0.3M, pH7.0).

Example 3: Production of antibodies Specific antibodies against the proteins according to the invention were produced. These were used to analyze the tissue distribution at the protein level of PARP2 and PARP3 by WESTERN immunoassay. Examples of production of said antibodies are indicated below. The following peptides were prepared by synthesis in a manner familiar to workers skilled in the production of antibodies. In some cases, a cysteine residue was attached to NOC terminal sequences to facilitate coupling to KLH (lame hemocyanin). PARP-2: NH2-MAARRRRSTGGGRARALNES-C02H (amino acids 1-20, SEQ ID NO: 23) NH2-KTELQSPEHPLDQHYRNLHC-C02H (amionoacids335-353 353; SEQ ID NO: 24) PARP-3: NH2-ckgrqagreedpfrstaealk-C02H (amino acids 25 -44 SEQ ID NO: 25) NH2-CKQQIARGFEALEALEEALK-C02H (amino acids 230- 248; SEQ ID NO: 26) - production of u? Anti-PARP3 antibody is described as a representative example. For human PARP3, polyclonal antibodies were activated in rabbits using a synthetic peptide having the peptide sequence H2N-KQQIARGFEALEALEALK-C02H (SEQ ID NO: 27) (amino acids 230-248 of the PARP3 protein sequence) ^^^^^^^^ ^ ^^^^ ^^^ & ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ ^^^^ ~. * Á? ~~ Í? J A? .1 human). The corresponding sequence of mice differs in this region only by one amino acid (H2N-KQQIARGFEALEALEEAMK-C02H; SEQ ID NO: 28). An N-terminal cysteine was also attached to enable the protein to be coupled to KLH. The rabbits were immunized a total of 5 times, at intervals of 7 to 14 days, with the KLH-peptide conjugate. The antiserum obtained was purified by affinity using the antigen. The specific IgG fraction was isolated from the serum using the respective peptides, for this purpose, they were initially immobilized in an affinity column, in a manner familiar to skilled workers. The respective antiserum was loaded onto this affinity column and the non-specific absorbed proteins were eluted with the buffer. The specifically bound IgG fraction was eluted with glycine buffer 0.2M HCL pH2.2. The pH was immediately elevated using a TRIS buffer 1M / HCL pH7.5. The eluate containing the IgG fraction was mixed 1: 1 (volume) with saturated ammonium sulfate solution and incubated at + 4 ° C for thirty minutes to complete the precipitation. The resulting precipitate was centrifuged at 10,000 g and, after removing the supernatant, dissolved in the minimum amount of PBS / TBS. The resulting solution was dialyzed against PBS / TBS in the ratio of 1: 100 (volume). The antibodies were adjusted to a concentration of approximately lOOμg of IgG / ML. PARP3 antibodies purified in this way had high specificity for PARP3. Considering that mouse PARP3 was well recognized, no cross reaction was observed cor. PARP1 or PARP2. Example 4: Analysis of Tissue Distribution by Immunoassay (Western blot) The tissue distribution at the protein level was also investigated for PARP2 and PARP3 by Western immunoassay. Preparation of mouse tissues for protein gels: Tissues or cells were homogenized using a Potter or Ultra-Turrax. "For this, 0.5 g of tissue (or cells) were incubated in 5 ml of buffer (TRIS-HCL lOmM pH7.5, lmM EDTA, 6mM MgC12), a protease inhibitor cocktail tablet (Boeringher Mannheim, No. Order: 1836153) and benzonase (grade of purity I, Merck) at 37 ° C for thirty minutes The samples of the mice were taken from the heart, lung, liver, spleen, kidneys, intestine, muscles, brain and for humans were taken from cells of kidney "embryonic (HEK293, kidney of human embryo). Protein Gels: The NuPAGE system provided by NOVEX was used in accordance with the instructions for protein gels. Polyacrylamide gels (NuPAGE 4-12% BisTris, NOVEX NP 0321), run buffer (MES-Running buffer, NOVEX) were used. ^^ ¡^^^^^ ^ ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ ^^^^^^ t ^^^^^^^^^^^^^^^^^ NP 0002), antioxidant (NOVEX NP 0005), protein size standard (Multi Mark Multi Colored Standard, NOVEX LC 5725 ), sample buffer (NuPAGE LDS Sample Buffer (4X), NOVEX NP 0007). The gels were run for 45 minutes at a voltage of 200V. Western analysis: Western blots were performed using the NOVEX System according to the instructions. A nitrocellulose membrane (Nitrocellulose pore size 45μm, NOVEX LC 2001) was used. The transfer took one hour at a current of 200 mA. The transfer buffer consisted of 50 ml of transfer buffer concentrate (NOVEX LP 0006), one ml of antioxidant (NOVEX NP 0002), 100 ml of analytical grade methanol and 849 ml of bidistilled water. In addition to the stains produced in this way, pre-stained spots were also used, for example from Chemicon (mouse brain stain, Chemicon, Catalog No.: NS 106 with tissues 1. Front bark, 2. Back bark, 3. Cerebellum, 4. Hippocampus, 5. Olfactory bulb, 6. Striatum, 7. Thalamus, 8. Medium brain, 9. Endonasal cortex, 10. Bridge, 11. Marrow, 12. Spinal column Antibody reaction with PARP3: Western spots were blocked in TBST (TBS + "Tween 20 0.3% with dry milk powder at 5% for about 2 hours (TBS: Tris pH 7.5 100 mM, 200 mM NaCl). The antibody reaction with the primary antibody (1: 1000 dilution) took place in TBST with 5% dry milk powder (see above) at room temperature for about 2 hours at 4 ° C throughout the night, with gentle shaking (vertical rotor). The process continued washing three times in TBST for 5 minutes. Incubation with the secondary antibody (anti-rabbit IgG, coupled to peroxidase, SIGMA A-6154, dilution 1: 2000) took place in TBST with 5% dry milk powder for 1 hour. Next, washing three times for 5 minutes each time, as above. Subsequent detection was based on chemiluminescence using the SUPER BLAZE kit (Pierce, Signal BLAZE Chemiluminiscent Substrate 34095) as established by the manufacturer. We used "Lumi-Film" (Chemiluminescent Detection Film, Boehringer Order No.: 1666916). The films were developed for approximately 2 min. (concentrated X-ray developer, ADEFO-Chemie GmbH), hydrated, fixed for approximately 4 min (Acidofix 85 g / 1 / AGFA), hydrated and then dried. Example 5: Preparation of Enzymes By comparison, 'human PARP1 was expressed with recombination in the baculovirus system in a manner familiar to partially purified skilled workers as described (Shah et al., Analytical Biochemistry 1995, 227, 1-13) . Bovine PARP1 in a purity of 30-50% (c = 0.22 mg / ml, specific activity 170 nmoles of ADP-ribose / min / mg • - ^ xtii üufc ^ L jgftga ^ gife ^ .. of total protein at 25 ° C) was obtained from BIOMOL (Order No. SE-165). Human and mouse PARP2 and PARP3 were expressed with recombination in the baculovirus system (Bac-toBac system, BRL Lifescience) For this purpose, the appropriate cDNAs were cloned into the vector pFASTBAC-1. of the recombinant baculovirus DNA by recombination in E.coli, was the transfection of the insect cells (Sf9 or "High-Five") with the appropriate recombinant baculovirus DNAs.

The expression of the corresponding proteins was verified with the Western blot analysis. The viral strains were amplified in a familiar way to the expert workers. Larger amounts of recombinant proteins were obtained by infecting 500 ml of the culture insect cell (2 x 10 'cells / ml) with virus in a MOT (multiplicity of infection, proportion of virus in relation to cells) of 5-10 and incubating for 3 to 4 days. The insect cells were transformed into pellet by centrifugation, and the proteins were purified from the pellet.

'Pellet. "The purification was carried out through classical methods of purification of proteins familiar to expert workers, detecting the enzymes with the appropriate specific antibodies. were also purified by affinity in a column of - ^ - j? YiBgiiM? Affinity of 3-amnobenzamide as described (Burtscher et al., Anal. Biochem 1986, 152: 285-290). The purity was > 90% Example -6: The assay systems to determine the activity of PARP2 and PARP3 and the inhibitory action of the effectors in PARP1, PARP2 and PARP3. a) The production of antibodies against poly (ADP-ribose) It is possible to use poly (ADP-ribose) as an antigen to generate anti-poly (ADP-ribose) antibody. The production of anti-poly (ADP-ribose) antibodies is described in the literature (Kanai Y. et al. (1974) Biochem Biophys Res Comm 59: 1, 300-306; Kawamaitsu H et al. (1984) Biochemistry 23, 3771-3777; Kanai Y et al. (1978) Immunlogy 34, 501-508). The following were used, among other materials: antibodies (polyclonal antiserum, rabbits) anti-poly (ADP-ribose), BIOMOL; Order No. SA-276, anti-poly (ADP-ribose) antibodies (monoclonal, mouse, clone 10H, hybridoma supernatant, purified by affinity). The antiserum or * monoclonal antibodies obtained from the supernatant hybridoma were purified by protein A affinity chromatography in a manner familiar to skilled workers, b) ELISA Materials: l ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ ^^^^^^^^^^ ^ ELISA color reagent: TMB mix, SIGMA T-8540 A 96-well microtiter plate (FALCON Micro-Test III ™ Flexible Test Plate, # 3912) was coated with histones ( SIGMA, H-7755). The histones, for this purpose, were dissolved in a carbonate buffer (Na2C03 0.05 M, pH 9.4) at a concentration of 50 μg / ml The individual wells of the microtiter plate were each incubated with 150 μl of this solution. histone at room temperature for about 2 hours or at 4 ° C overnight The cavities are blocked, then, adding 150 μl of a 1% BSA solution (SIGMA, A-7888) in carbonate 'buffer' at room temperature environment for 2 hours. Next, three washing steps are carried out with washing 'buffer' (Tween 10 0.05% in 1 x PBS, PBS (Saline 'buffered' with phosphate, Gibco, No. of order 10010): KH; P04 0.21 g / 1, NaCl 9g / l, NaHPO.¡.7H: 0 0.726 g / 1, pH 7.4). The washing steps were carried out in a microtiter plate washer ('Columbus' microtiter plate washer, SLT-Labinstruments, Austria) For the enzymatic reaction an enzyme reaction solution and a substrate solution were used, in each case as A premix The absolute amount of these solutions depends on the test cavities. ..4 k 'Ski? - go, ... ____ ^^^^ _ ^^^^^^^^^^^^^^^^^^ >; h ^ Composition of the enzyme reaction solution per cavity: 4 μl of PARP reaction buffer (Tris-HCl IM, pH 8.0, MgCl, 100mM, 10mM DDT) - 20 ng of PARP1 (human or bovine) or 8 ng of PARP2 (human or mouse) 4 μl of activated DNA (1 mg / ml, SIGMA, D-4522) H20 at 40 μl The composition of the substrate solution per well: - 5 μl of reaction buffer PARP (lOx) 0.8 μl of NAD solution (10 mM, SIGMA N-1511) 44 μl of H20 The inhibitors were dissolved in the PARP lx reaction buffer, DMSO, which was occasionally used to dissolve the inhibitors at higher concentrations, did not present problems at a final concentration of 2% For the enzymatic reaction, 40 μl of enzyme reaction solution was introduced into each of the cavities and se1 incubated with 10 μl of inhibitory solution for 10"minutes. The enzymatic reaction was started, then, by adding 50 μl of substrate solution per cavity. The reaction was carried out at room temperature for 30 minutes and then extinguished by washing three times with washing buffer. ^^ l ^^^^^^^^^^^ ^^ gj ^^^^ xj ^^^^? ^^^^^^^^^^ g ^ j ^^^^^^ t ^^^ ^ ^ ^ ^^^^^^^^^ J ^^^^ The antibodies used were specific anti-poly (ADP-ribose) antibodies at a dilution of 1: 5000. Dilution was carried out in antibody buffer (1% BSA in PBS, 0.05% Tween 20) The incubation time for the primary antibodies was one hour at room temperature, after subsequent washing three times with 'buffer' After washing, the incubation was carried out with the anti-mouse IgG secondary antibody, Gab fragments, coupled to the peroxidase, Boehringer Mannheim, No. of order 1500.686; IgG enticonejo, coupled to peroxidase, SIGMA, No. of order A-6154) in a dilution of 1: 10000 in 'buffer' of antibodies at room temperature for one hour After washing three times with 'buffer' of washing, performed a color reaction using 100 μl of color reagent (TMB mixture, SIGMA) per well at room temperature for about 15 minutes. The color reaction was terminated by adding lOOμl of 2M H2SO4. It was then measured immediately on an ELISA plate reader (EAR340AT 'Easy Reader', SLT-Labisntruments, Austria) (450 nm against 620 nm) The principle of measurement is described in diagram form in Figure 6. Various concentrations were used to construct a dose-effect diagram to determine the Ki value of an inhibitor.The values were obtained in triplicate for a particular concentration of the inhibitor. f ^ ^ j __ ^^^ j 'arithmetics were determined using Microsoft © Excel. The IC = .- is determined using the Microcal Software © Origin (version 5.0) ('Sigmoidal Adjustment'). The conversion of the value IC5j to Ki values which is calculated in this way, was done using 'calibration inhibitors'. The 'calibration inhibitors' were also measured in each analysis.The K-values of the calibration inhibitors were determined in the same test system by analysis of the Dixon Diagram in a manner familiar to the skilled workers b) HTRF assay (Homogeneous resolution time fluorescence) In the PARP assay by HTRF according to the invention, the histones, as target proteins that will be modified by PARP, are labeled indirectly with an XL665 fluorophore.The anti-poly (ADP-ribose) antibody it is directly labeled with a europium cryptate (anti-PAR-cryptate) If the fluorophore is in the direct neighborhood in space, which is secured by binding to the poly (ADP-ribose) in the histone, the energy transfer is possible The emission at 665 nm is, therefore, directly proportional to the amount of bound antibodies, which in turn is equivalent to the amount of poly (ADP-ribose) .The measured signal of that form corresponds to the active PARP principle The measurement principle is described in diagram form in Figure 7. The materials used are ? j ^ i ^ i ^^^ egj j ^^^! ^ 3jáf s ^^ ^^^^^ ^^^^ j ^ j ^ identical to those used in the ELISA (see above), unless be expressly indicated. The histones were dissolved in a concentration of 3 mg / ml in Hepes buffer (50 mM, pH 7.5) Biotinylation was performed with 5 sulfo-NHS-LC-biotin (Pierce, # 21335T). of 4 biotin molecules per histone The incubation time was 90 minutes (RT) The biotinylated histones were purified on a G25 SF HR10 / 10 column (Pharmacia, 17-0591-01) in Hepes buffer (50mM, pH 7.0) or to remove excess biotinylation reagent. The anti-poly (ADP-ribose) antibody was labeled with europium cryptate using bifunctional coupling reagents (Lopez, E. et al., Clin. Chem. 39 (2), 196-201 (1993); US Patent 5,534,662). purification was performed in a! 5 column G25 SF HR10 / 30. The molar ratio of 3.1 of cryptates per antibody was reached. The yield was 25%. The conjugates were stored at -80 ° C in the presence of 0.1% BSA in phosphate buffer (0.1M, pH = 7.) For the enzymatic reaction, the following compositions were pipetted into each of the cavities: lOμl of solution of PARP in PARP HTRF reaction buffer (50 mM Tris-HCl pH 8.0, 10 mM MgCl, 1 mM DTT) with 20 ng of PARP1 (human or bovine) or 8 ng of PARP2 (human or mouse) 25 lOμl of DNA activated in HTRF reaction buffer PARP (50 μl / ml) lOμl of biotinylated histones in HTRF reaction buffer of PARP (1.25 μM) - lOμl of "inhibitor in HTRF reaction buffer of PARP The reagents were incubated for 2 minutes before starting the reaction with the addition of 10 μL of NAD solution in HTRF reaction buffer.

PARP (41μM / ml). The reaction time was 30 minutes at room temperature. The reaction was inhibited by adding 10 μl of PARP inhibitor (25 μM, Ki = lOnM) in 'revelation' buffer (Tris-HCl lOOmM pH 7.2, 0.2M KF, BSA 0. 05%). Then the following was added: lOμl of EDTA solution (SIGMA, E-7889, 0.5M in water) lOOμl of Sa-XL665 (Packard Instruments) in buffer 'Revelation' (15-31.25nM) 50μl of anti-PAR cryptate in 'Revealed' buffer (1.6-3.3 nM). The measurements could be made after 30 minutes (up to 4 hours). The measurements were made on a 'discovery HTRF microplate analyzer' (Canberra Packard Instruments) The kx values were calculated as described for the ELISA Example 7: Test Systems to determine the therapeutic efficiency of the PARP inhibitors. The therapeutic efficiency of the new PARP inhibitors in relevant pharmacological models Examples of some suitable models are presented in Table 1. fifteen twenty ^ J ^^ g ^^^ tójtó ^ s * ^^^^^^^^ - rats or mice 1997, 3: 1089-1095. Endres, M et al., J Cereb Blood Flow Metab 1997, 17: 1143-1151. Takashi K et al., J Cereb Blood Flow Metab 1997, 17: 1137-1142. 10 Toxicity Disease of MPTPlCosi C, and others, Parkinson (1-methyl-4-phenyl-Brain Res., 1998 1,2,3,6-809 (1): 58-67, tetrahydropyridine) COSÍ C, and others , in mice / rats Brain Res., 1996 729 (2): 264-9. Infarction of vessel occlusion Richard V, and others; myocardium in rats, pigs Br. J. Pharmacol or rabbits 1994, 113, 869-876. Thiemermann, C, and others, Proc Natl Acad Sci, USA. 1997, 94 (2): 679-83. Zingarelli B. Y ^ ¡¡^^^ ¿^ ¡¡^^^^^ ¡^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ "" "< ** • * - * • < ^^^ ¡^^^^^^^^ x Diabetologia 1985, 28: 683-686. Shimabukuro M and 1 others, J Clin Invest 1997, 100: 290-295. Cancer In vitro model; Schlicker et al., See below 1999, 75 (1): 91-100 a) NMDA excitotoxicity model Glutamate is the most important neurotransmitter in the brain. Under normal conditions, glutamate is secreted in synaptic clefts and stimulates post-synaptic glutamate receptors, specifically glutamate receptors of the 'NMDA' and 'AMPA' type. Stimulation plays a significant part in numerous brain functions, including learning, memory and motor control. Under conditions of chronic and acute neurodegeneration (for example, stroke), however, there is a large increase in the secretion of pre-synaptic glutamate, resulting in excessive stimulation of the receptors. This leads to the death of cells stimulated in this way.

These increased glutamate activities occur in a number of neurological conditions or physiological disturbances and lead to states of overexitation or toxic effects in the central nervous system (CNS), but also in the peripheral nervous system. In this way, glutamate is involved in a large number of "neurodegenerative conditions, in particular neurotoxic disorders that follow hypoxia, anoxia, ischemia and after injuries such as those that occur after stroke and trauma, Alzheimer's disease, of Huntington, amyotrophic lateral sclerosis (ALS, Lou Gehring's disease), cranial trauma, spinal cord trauma, peripheral neuropathies, AIDS dementia and Parkinson's disease Another disease in which glutamate receptors are important is epilepsy (cf. Brain Res Bull 1998; 46 (4): 281-309, Eur Neuropsychophar acol 1998, 8 (2): 141-52.) The effects of glutamate are mediated through several receptors One of these receptors is called NMDA receptor (N-methyl-D-aspartate) after a specific agonist (Arzneim, Forschung 1990, 40, 511-514).; TIPS, 1990, 11, 334-338; Drugs of the Future 1989, 14, 1059-1071). N-methyl-D-aspartate is a strong agonist of a particular class of glutamate receptors (type 'NMDA'). NMDA receptor stimulation leads to the entry of calcium in the cell and the generation of free radicals.

~ Ji-a '- "- *' '•" * i f f »itnt« r-f hM * Uai_ftM? ________________.

Free radicals lead to DNA damage and activation of PARP. PARP, in turn, causes cell death through the decrease of high energy phosphates (NAD and ATP) in the cell. This explains the toxicity of NMDA. The treatment of animals with NMDA can, therefore, be taken as a model of the aforementioned conditions in which excitotoxicity is involved. Due to the importance of glutamate receptors in neurodegeneration, many pharmacological approaches to block specific or precisely these receptors have been directed to date. However, due to their importance in the normal conduction of the stimuli, these approaches were problematic (collateral effects). Additionally, the stimulation of the receivers is an event that takes place very quickly, so that the administration of the receivers in general comes very late (problem of the 'time window') .There is therefore a great need for new Principles of action and inhibitors of NMDA-related neurotoxicity Protection against brain over-excitation by excitatory amino acids (NMDA antagonism in mice) can be seen as an adequate test of the activity of a pharmacological effector of PARP in conditions based on excitotoxicity. Intracerebral administration of excitatory amino acids (EEA) induces such overexcitation ^^^^^^^^^^^^^^^^^^^^ JS ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ time to convulsions and death of animals (mice). In the present case, 10 μl of an aqueous NMDA solution of force 0.035% intracerebroventricularly was administered unilaterally, 120 minutes after the intraperitoneal administration (i.p.) of the tested substance. These symptoms can be inhibited by systemic administration, for example, intraperitoneal, of drugs that act centrally. Because excessive activation of the EEA receptors in? Or the central nervous system plays an important role in the pathogenesis of several neurological disturbances, information on the EEA antagonism detected in vivo can be obtained by possible therapeutic use of the substances in such conditions. of the SNC. It was determined as a measure of activity of the substances, an ED50 in which 50% of the animals are free of symptoms with a fixed dose of NMDA, due to the previous intraperitoneal administration of the substance measured, b) Langendorf heart model (model for infarction of myocardium) Male Sprague-Dawley rats (body weight, 300-400 g, origin: Janvier, Le Genest-St-Isle, France) were used for the test. The rats were treated orally by priming with the active substance or placebo (volume: 5 ml / kg). 50 minutes then, heparin was administered intraperitoneally (Liquemin N Roche, 125 IU / animal in 0.5 ml). The animals were anesthetized with Inactin® T133 (10% sodium thiobetabarbital), fixed on the operating table, underwent a tracheotomy and ventilated with a 'Harvard ventilation pump' (40 beats / minute, 4.5 ml / beat The thoracotomy was performed, followed by immediate catheterization of the aorta, removal of the heart and immediate retrograde perfusion.The hearts were perfused with a constant pressure of 75 mmHg, which was achieved using a 'perfusion pump 2 Gilson. Miniplus. " The perfusate composition (mmol / L): NaCl 118, KCl 4.7, CaCl; x 2 H: 02.52, MgSO- x 7 H20 1.64, NaHCOs 24.88, KH: PO 1.18, glucose 11. The temperature was maintained at 37 ° C throughout the experiment. Functional parameters were recorded continuously using a '5-channel Gould recorder. "The measures that were performed were: left ventricular pressure (LVP; mmHg), LVEDP (mmHg), enzymatic release (creatine kinase, mU / ml / g) , coronary flow velocity (ml / min), HR (pulse rate, min) Left ventricular pressure was measured using a latex balloon filled with fluid and a Stataham 23 Db pressure transducer. The volume of the balloon was initially adjusted to achieve a LVEDP (left ventricular end diastolic pressure) of approximately 12 mmHg The dP / dtma (maximum pumping force) is derived from the pressure signal using a different module. j ^^^^^^ ijj ^ ktí ^ _ ^^ jM «^^) ^^ j» g ^^^? ^ jj ^^ j ^^? fc * j 4áíi ^^ A ¿Cardiac system was determined drip counter (BMT Messttechnick GmbH Berlin). After a balance time of 20 minutes, the hearts underwent a 30-minute global ischemia, stopping the perfusate supply, while maintaining the temperature at 37 ° C. During the next 60 minutes of perfusion period, the perfusate samples were taken after 3, 5, 10, 15, 30, 45 and 60 minutes for the analysis of creatine kinase (CK) activity. The averages and standard deviations for the measured parameters were analyzed statistically (Dunett's test). The significant limit was p = 0.05.

The experiment in rabbit hearts was carried out similarly. Male New Zealand white rabbits (obtained from Interfauna) were used. The hearts were prepared as described above for the rat model. The perfusion pressure was set at a maximum of 60 mmHg and the flow rate at approximately 25 ml / min. The equilibrium time was approximately 30 minutes. The substance was administered by infusion directly ascending in relation to the heart. 15 minutes after the infusion started, a global ischemia was provoked for 30 minutes stopping the flow, while the heart temperature was maintained. The procedure followed with a perfusion of 30 minutes. The perfusate was taken to investigate the activity of CK before the administration of the substance, after 15 minutes and at various times during the perfusion (5, 10, 15, 20 and 30 minutes). The following parameters were measured: LVP (mmHg), LVDP, LVdp / dt, PP (mmHg), HR (pulse rate, beats / min), CK activity (U / min / g of heart weight). c) Animal model for acute kidney failure The protective effect of intravenous administration of PARP inhibitors (4 days) on kidney function in rats with acute kidney failure post-ischemic was investigated. Male Sprague-Dawley rats of approximately 330 g were used at the start of the experiment; breeder: Charles River). 10-15 animals were used per experimental group. The administration of the active substance / placebo was performed continuously with an osmotic micropump in the femoral vein. Orbital blood was taken (1.5 ml of whole blood) under inhalation anesthesia with eflurane (Ethrane Abbot, Weisabaden). After the initial measurements (blood samples) and determination of the amount of urine excreted in 24 h, the rats were anesthetized ('Nembutal', sodium pentobarbital, Sanofi CEVA, 50 mg / kg ip, volume injected 1.0 ml / kg) and were fixed on a heated operating table (37 ° C), 125 IU / kg of heparin (Liquemin N, Roche) were administered intravenously into the caudal vein. ^ ^ ^ ^ ^ ^ ^ ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ ^^^^^^ gg ^ ¡^ ¡^^^^ i ^^ abdominal and exposed the right kidney. The branch of the renal artery was exposed and grasped on its upper part using bulldog fasteners (Diefenbach 38 mm). The left renal artery was exposed in a similar way and was held (in its upper part 5, approximately halfway to the kidney). During the operation, "an osmotic micropump was implanted in the femoral vein, the intestine was reinserted and the fluid loss was compensated with warm 0.9% NaCl. The animals were covered with a damp cloth and kept warm. After 40 minutes, the appearance of the kidneys was checked, and the fasteners were removed, first the right and then the left, the intestine was replaced and two drops of antibiotic were added (Tardomyocel, Bayer). abdominal was closed with sterilized cat intestine (Ethicon No. 4) and treated once more with 1 drop of antibiotic. The epidermis was sutured with Ethibond Exel (Ethicon) sterile No.3 / 0, and the suture was sprayed with wound spray Nebacetin N (Yamanouchi). One tenth of a daily dose of drug / placebo is applied as a bolus intravenously. Samples and blood were taken to investigate biochemical parameters in serum and urine: Na, K,. Creatinine, proteins (only in urine), on days 1, 2 and 4 of the experiment. In addition, the consumption of food and water, the weight was recorded * ~ ** am. * aa. *** > . . rt ..- ¿¡. ", > .. S,.,: > < , - ". -, - »". _ * __ .. _. body and the volume of urine. 14 days later, the animals were sacrificed and the kidneys were evaluated. The evaluation excluded all animals that died due to infarction during the experiment or showed infarction at the 14th necropsy. Creatinine clearance and fractional sodium excretion were calculated as parameters of renal function, comparing the treated and control animals. simulation, d) In vitro model for radiosensis bilization (tumor therapy) MCF-7 (human mammary carcinoma) cells were cultured in a modified Dulbecco's Eagle medium with 10% heat-inactivated FCS and 2mM L-glutamine. Cells were seeded overnight at cell densities of 100, 1000 or 10000 cells per well in a 6-well plate and then exposed to ionizing radiation at a dose in the range of 0 to 10 Gy (1J "" CS, Shepard Mark, model I-68A, dose intensity 3.28 Gy / min). 10 days after the irradiation, the experiment was evaluated, considering the colonies positive with fifty cells. e) Cerebral Stroke Model (focal cerebral ischaemia, middle cerebral artery occlusion (MCA) in a rat) Ischemia was caused by cauterization of the right distal MCA in Sprague-Dawley rats or Long-Evans rats. Rats can be treated before or after the start of ^^^ and ^^ ¿¿^^^^ j ^ g ^^^^ g ^ ^^^^^ J ^^ gl ^ | ^^^^ ^ ^ g ^ ^ gi ^ ^^^^ _ ^^ ^ __ ^^ _ ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ As a rule, doses of 1-10 mg / kg (application of a bolus) are chosen, optionally followed by a continuous infusion of 0.5-5 mg / kg / h. The rats are anesthetized with halothane in a mixture of 70% nitrogen and 30% oxygen (4% 'in the initial fasse and 0.8-1.2% during the operation). The body temperature was permanently measured rectally and kept constant at 37.5 ° C ± 0.5 ° C by means of an adjustable heating blanket. Moreover, arterial blood pressure, pH, Pa (02) and Pa (C02) were optionally measured by means of a tail venous catheter. Subsequently, focal ischemia occurred using the method of Chen et al. (Stroke 17: 738-743; 1986) or Liu et al. (AmJ Physiol 256: H589-593; 1989) by continuous cauterization of the distal part of the right AMC. When the operation was finished, the animals were kept in a warm environment for another 24 hours. They were then slaughtered using CO: and decapitated. The brains were extracted, frozen instantly (with dry ice or liquid nitrogen) and kept at -80 ° C. The brains were cut into 0.02 mm thick sheets and one in 20 was used for further analysis. The corresponding cuts are. stain with cresyl violet (Nissl stain). As an alternative, TTC (2, 3, 4-triphenyltetrazolium chloride) can be used • üßbÉiÉi ri for staining. The infarct volume can then be analyzed under a microscope. For an exact quantification, a computer image analysis program (J Cereb Blood Flow Metabol 10: 290-293, 1990) can be used. f) Septic Shock Groups of 10 male C57? BL mice (body weight 18-20 g) are treated with LPS (lipopolysaccharide from E. coli, LD100 20 mg / animal, intravenous foram) plus galactosamine (20 mg / animal, in endovenous form). The substance to be tested is applied intraperitoneally or intravenously for three consecutive days (eg 1-10 mg / kg), the first dose being administered 30 minutes after treatment with LPS. The death rate is determined every 12 hours. Alternatively, the substance can also be applied in several doses distributed on those days. g) Determination of altered gene expression in cells with aging Cell aging is simulated by changing the culture media of the cells from a complete medium to a medium with reduced serum concentration and subsequently analyzed by means of quantitative PCR or by Northern immunoassay (Linskens et al., Nucleic Acids Res. 1995: 23 (16): 3244-51). As typical markers for skin aging, for example, collagen or elastin can be used. Fibroblasts or fibroblast cell lines are used ^^ '^^ i jij ^^^^^^ W ^ yj ^^^ ^^^ lßíáí g * »^^^^^^^^^^^^ human that simulate the aging p. Modulators of the proteins of the invention are added to the medium and their effect on the change of gene expression is observed. An increased production of elastin can be seen in cells with a reduced aging process caused by said modulators. 0 LIST OF SEQUENCES (1) GENERAL INFORMATION (i) APPLICANT: • (A) NAME: BASF Ahtiengesellschaft (B) ADDRESS: (C) CITY: Ludwigshafen (E) COUNTRY: Germany (F) POSTAL CODE: 67065 (ii) TITLE OF THE INVENTION: New Generation of ADP Ribosa Poii erasa (iii) SEQUENCE NUMBER: 28 (iv) LEGIBLE FORM FOR THE COMPUTER: (A) TYPE OF MEDIA: Floppy Disk (B) COMPUTER: IBM compatible PC (C) OPERATING SYSTEM: PC DOS / MS DOS (D) SOFTWARE: Patentln Relay # 1.0, Version # 1.30 (EPO) (2) INFORMATION FOR SEQ ID NO: 1: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 1843 base pairs (B) TYPE : Nucleic acid (C) TYPE OF HEBRA: Monohebra (D) TOPOLOGY: Linear (ii) TYPE OF MOLECULE: Cdna (ni) HYPOTHETICAL: No (iv) ANTIPARALELO: No (vi) ORIGINAL SOURCE: (E) TISSUE UNCLE: Brain (ix) CHARACTERISTICS: (A) NAME / KEY: CDS (B) LOCATION: 3..1715 (C) OTHER INFORMATION: / pipeline = "poly ADP Ribose Polymerase" (Xi) DESCRIPCION.DE SEQUENCE: SEQ ID NO: 1: CC ATG CGA GCG GCG CGG CGG CGG ACC AGC GGC GGC GGC GCG AGG AGA 47 Met Ala Ala Arg Arg Arg Arg Ser Thr Gly Gly Gl? Ala Arg Arg1 May 10 15 GCA TTA AGC AAT AAA GAA AGA GTT AAT AAC AAT GGC ACG CCA GCT GAA 95 Ala Leu Asn Glu Ser Lys Arg Val Asn Asn Gly Asn Thr Ala Pro Glu 20 25 30 GAC TCT TCC CCT GCC AAG AAA ACT CGT AGA TGC CAG AGA CAG GAG TCG 143 Asp Ser Ser Pro Ala Lys Lys Thr Arg Arg Cys Gln Acg Gln Glu Ser 35 40"45 ** & * & * *: AAA AAG ATG CCT GTG GCT GGA GGA AAA GCT AAT AAG GAC AGG ACA GAA 191 Lys Lys Met Pro Val Wing Gly Gly Lys Wing Asn Lys Asp Arg Thr Glu 50 55 60 GAC AAG CAA GAT GAA TCT GTG AAG GCC TTG CTG TTA AAG GGC AAA GCT 239 Asp Lys Gln Asp Glu Ser Val Lys Ala Leu Leu Leu Lys Gly Lys Wing 65 70 75 CCT GTG GAC CCA GAG TGT ACÁ GCC AAG GTG GGG AAG GCT CAT GTG TAT 287 Pro Val Asp Pro Glu Cys Thr Ala Lys Val Gly Lys Ala His Val Tyr 80 85, 90 95 TGT GAA GGA AAT GAT GTC TAT GAT GTC ATG CTA AAT CAG ACC AAT CTC 335 Cys Glu Gly Asn Asp Val Tyr Asp Val Met Leu Asn Gln Thr Asn Leu 100 105 110 CAG TTC AAC AAC AAC AAG TAC TAT CTG ATT CAG CTA TTA GAA GAT GAT 383 Gln Phe Asn Asn Asn Lys Tyr Tyr Leu lie Gln Leu Leu Glu Asp Asp 115 120 125 GCC CAG AGG AAC TTC AGT GTT TGG ATG AGA TGG GGC CGA GTT GGG AAA 431 Wing Gln Arg Asn Phe Ser Val Trp Met Arg Trp Gly Arg Val Gly Lys 10 130 135 140 ATG GGA CAG CAC AGC CTG GTG GCT TGT TCA GGC AAT CTC AAC AAG GCC 479 Met Gly Gln His Ser Leu Val Ala Cys Ser Gly Ace n Leu Asn Lys Wing 145 150 155 AAG GAA ATC TTT CAG AAG AAA TTC CTT GAC AAA ACG AAA AAC AAT TGG 527 Lys Glu lie Phe Gln Lys Lys Phe Leu Asp Lys Thr Lys Asn Asn Trp 160 165 170 175 GAA GAT CGA GAA AAG TTT GAG AAG GTG CCT GGA AAA TAT GAT ATG CTA 575 5 Glu Asp Arg Glu Lys Phe Glu Lys Val Pro Gly Lys Tyr Asp Met Leu 180 185 190 CAG ATG GAC TAT GCC ACC AAT ACT CAG GAT GAA GAG GAA ACA AAA 623 Gln Met Asp Tyr Ala Thr Asn Thr Gln Asp Glu Glu Glu Thr Lys Lys 195 200 205 GAG GAA TCT CTT AAA TCT CCC TTG AAG CCA GAG TCA CAG CTA GAT CTT 671 Glu Glu Ser Leu Lys Ser Pro Leu Lys Pro Glu Ser Gln Leu Asp Leu 210 215 • 220 CGG GTA CAG GAG TTA ATA AAG TTG ATC TGT AAT GTT CAG GCC ATG GAA 719 20 Arg Val Gln Glu Leu lie Lys Leu lie Cys Asn Val Gln Wing Met Glu 225 230 235 'GAA ATG ATG ATG GAA ATG AAG TAT AAT ACC AAG AAA GCC CCA CTT GGG 767 Glu Met Met Met Glu Met Lys Tyr Asn Thr Lys Lys Ala Pro Leu Gly 240 245 250 255 AAG CTG ACÁ GTG GCA CAA ATC AAG GCA GGT TAC CAG TCT CTT AAG AAG 815 Lys Leu Thr Val Wing Gln lie Lys Wing Gly Tyr Gln Ser Leu Lys Lys 260 265 270 - «a»? ^ TB¡ * ¡.b «¡.. ... -r i ... A ... .. ...,. . ... . ..... .. . ...,. ... ... .. AHA......

ATT GAG GAT TGT ATT CGG GCT GGC CAG CAT GGA CGA GCT CTC ATG GAA 863 He Glu Asp Cys He Arg Wing Gly Gln His Gly Arg Wing Leu Met Glu 275 280 285 GCA TGC AAT GAA TTC TAC ACC AGG ATT CCG CAT GAC TTT GGA CTC CGT 911 Ala Cys Asn Glu Phe Tyr Thr Arg He Pro His Asp Phe Gly Leu Arg 290 295 300 ACT CCT CCA CTA ATC CGG ACÁ CAG AAG GAA CTG TCA GAA AAA ATA CAÁ 959 Thr Pro Pro Leu He Arg Thr Gln Lys Glu Leu Ser Glu Lys He Gln 305 310 315 TTA CTA GAG GCT TTG GGA ATT GAA ATT GCT ATT AAG CTG GTG AAA 1007 Leu Leu Glu Ala Leu Gly Asp He Glu He Wing He Lys Leu Val Lys 320 325 330 335 ACÁ GAG CTA CAA AGC CCA GAA CAC CCA TTG GAC CAÁ CAC TAT AGA AAC 1055 Thr Glu Leu Gln Pro Pro Glu His Pro Leu Asp Gln His Tyr Arg Asn 340 345 350 CTA CAT TGT GCC TTG CGC CCC CTT GAC CAT GAA AGT TAC GAG TTC AAA 1103 Leu His Cys Ala Leu Arg Pro Leu Asp His Glu Ser Tyr Glu Phe Lys 355 360 365 GTG ATT TCC CAG TAC CTA CAA TCT ACC CAT GCT CCC ACA CAC AGC GAC 1151 Val He Ser Gln Tyr Leu Gln Ser Thr His Wing Pro Thr His Ser Asp 370 375 380 TAT ACC ATG ACC TTG CTG GAT TTG TTT GAA GTG GAG AAG GAT GGT GAG 1199 Tyr Thr Met Thr Leu Leu Asp Leu Phe Glu Val Glu Lys Asp Gly Glu 385 390 395 AAA GAA GCC TTC AGA GAG GAC CTT CAT AAC AGG ATG CTT CTA TGG CAT 1247 Lys Glu Wing Phe Arg Glu Asp Leu His Asn Arg Met Leu Leu Trp His 400 405 410 415 GGT TCC AGG ATG AGT AAC TGG GTG GGA ATC TTG AGC CAT GGG CTT CGA 1295 Gly Ser Arg Met Ser Asn Trp Val Gly He Leu Ser His Gly Leu Arg 420 425 430 ATT GCC CCA CCT GAA GCT CCC ATC ACÁ GGT TAC ATG TTT GGG AAA GGA 1343 He Wing Pro Pro Glu Wing Pro He Thr Gly Tyr Met Phe Gly Lys Gly 435 440 445 ATC TAC TTT GCT GAC ATG TCT TCC AAG AGT GCC AAT TAC TGC TTT GCC 1391 He Tyr Phe Wing Asp Met Ser Ser Lys Ser Wing Asn Tyr Cys Phe Wing 450 455 460 TCT CGC CTA AAG AAT ACÁ GGA CTG CTG CTC TTA TCA GAG GTA GCT CTA 1439 Being Arg Leu Lys Asn Thr Gly Leu Leu Leu Leu Ser Glu Val Wing Leu 465 470 475 GGT CAG TGT AAT GAA CTA CTA GAG GCC AAT CCT AAG GCC GAA GGA TTG 1487 Gly Gln Cys Asn Glu Leu Leu Glu Wing Asn Pro Lys Wing Glu Gly Leu ^^^^^^^^^^^^^^^^^^^^^ 4¿ ^^^ * a ^^^ 6 ^^? ^^ < ^^^^ a ^ ití ^^^^^^^^^^^^ ^ ^ ^ ^ ^ ^ ^ 480 485 490 495 CTT CAÁ GGT AAA CAT AGC ACC AAG GGG CTG GGC AAG ATG GCT CCC AGT 1535 Leu Gln Gly Lys His Ser Thr Lys Gly Leu Gly Lys Met Wing Pro Ser 500 505 510 TCT GCC CAC TTC GTC ACC CTG AAT GGG AGT ACÁ GTG CCA TTA GGA CCA 1583 Ser Ala His Phe Val Thr Leu Asn Gly Ser Thr Val Pro Leu Gly Pro 515 520 525 GCA AGT GAC ACA GGA ATT CTG AAT CCA GAT GGT TAT ACC CTC AAC TAC 1631 Wing Ser Asp Thr Gly He Leu Asn Pro Asp Gly Tyr Thr Leu Asn Tyr 530 535-540 AAT GAA TAT ATT GTA TAT AAC CCC AAC CAG GTC CGT ATG CGG TAC CTT 1679 Asn Glu Tyr He Val Tyr Asn Pro Asn Gln Val Arg Met Arg Tyr Leu 545 550 555 TTA AAG GTT CAG TTT AAT TTC CTT CAG CTG TGG TGA ATGTTGATAT 1725 Leu Lys Val Gln Phe Asn Phe Leu Gln Leu Trp * 560 565 570 TAAATAAACC AGAGATCTGA TCTTCAAGCA AGAAAATAAG CAGTGTTGTA CTTGTGAATT 1785 TTGTGATATT TTATGTAATA AAAACTGTAC AGGTCTAAAA AAAAAAAAAA AAAAAAAA 1843 (2) INFORMATION FOR SEQ ID NO: 2: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 571 amino acids (B) TYPE: amino acid (D) TOPOLOGY: Linear (ii) TYPE OF MOLECULE: Protein (xi) DESCRIPTION FROM THE SEQUENCE: SEQ ID NO: 2: Met Ala Ala Arg Arg Arg Arg Ser Thr Gly Gly Arg Ala Arg Ala 1 5 10 15 Leu Asn Glu Ser Lys Arg Val Asn Asn Gly Asn Thr Ala Pro Glu Asp 20 25 30 Ser Ser Pro Ala Lys Lys Thr Arg Arg Cys Gln Arg Gln Glu Ser Lys 35 40 45 Lys Met Pro Val Wing Gly Gly Lys Wing Asn Lys Asp Arg Thr Glu Asp 50 55 • 60 Lys Gln Asp Glu Ser Val Lys Ala Leu Leu Leu Lys Gly Lys Wing Pro 65 70 75 80 Val Asp Pro Glu Cys Thr Wing Lys Val Gly Lys Wing His Val Tyr Cys 85 90 95 Glu Gly Asn Asp Val Tyr Asp Val Met Leu Asn Gln Thr Asn Leu Gln 100 105 110 Phe Asn Asn Asn Lys Tyr Tyr Leu He Gln Leu Leu Glu Asp Asp Wing 115 120 125 ^ Üfe ^^ Gln Arg Asn Phe Ser Val Trp Met Arg Trp Gly Arg Val Gly Lys Met 130 135 140 Gly Gln His Ser Leu Val Wing Cys Ser Gly Asn Leu Asn Lys Wing Lys 145 150 155 160 Glu He Phe Gln Lys Lys Phe Leu Asp Lys Thr Lys Asn Asn Trp Glu 165 170 »175 Asp Arg Glu Lys Phe Glu Lys Val Pro Gly Lys Tyr Asp Met Leu Gln 180 185 190 Met Met Asp Tyr Ala Thr Asn Thr Gln Asp Glu Glu Glu Thr Lys Lys Glu 195 200 205 Glu Ser Leu Lys Ser Pro Leu Lys Pro Glu Ser Gln Leu Asp Leu Arg 210 215 220 Val Gln Glu Leu He Lys Leu He Cys Asn Val Gln Ala Met Glu Glu 225 230 235 240 Met Met Met Glu Met Lys Tyr Asn Thr Lys Lys Wing Pro Leu Gly Lys 245 250 255 Leu Thr Val Wing Gln He Lys Wing Gly Tyr Gln Ser Leu Lys Lys He 260 265 270 Glu Asp Cys He Arg Wing Gly Gln His Gly Arg Wing Leu Met Glu Wing 275 280 285 Cys Asn Glu Phe Tyr Thr Arg He Pro His Asp Phe Gly Leu Arg Thr 290 295 300 15 Pro Pro Leu He Arg Thr Gln Lys Glu Leu Ser Glu Lys He Gln Leu 305 310 315 320 Leu Glu Wing Leu Gly Asp He Glu He Wing He Lys Leu Val Lys Thr 325 330 335 Glu Leu Gln Ser Pro Glu His Pro Leu Asp Gln His Tyr Arg Asn Leu 340 345 350 His Cys Ala Leu Arg Pro Leu Asp His "Glu Ser Tyr Glu Phe Lys Val 355 360 365 20 He Ser Gln Tyr Leu Gln Ser Thr His Wing Pro Thr His Ser Asp Tyr 370 375 380 Thr Met Thr Leu Leu Asp Leu Phe Glu Val Glu Lys Asp Gly Glu Lys 385 390 395 400 Glu Ala Phe Arg Glu Asp Leu His Asn Arg Met Leu Leu Trp His Gly 405 410 415 Ser Arg Met Ser Asn Trp Val Gly He Leu Ser His Gly Leu Arg He 420 425.430 25 Wing Pro Pro Glu Wing Pro He Thr Gly Tyr Met Phe Gly Ly s Gly He ^ k¡¡¡ttmmJái MM ^^ 435 440 445 ^ Tyr Phe Ala Asp Met Ser Ser Lys Ser Wing Asn Tyr Cys Phe a Ser 450 955 460 Arg Leu Lys Asn Thr Gly Leu Leu Leu Leu Ser Glu Val Ala Leu Gly 465 470 475 480 Gln Cys Asn Glu Leu Leu Glu Wing Asn Pro Lys Wing Glu Gly Leu Leu 485 490 495 Gln Gly Lys His Ser Thr Lys Gly Leu Gly Lys Met Wing Pro Ser Ser 500 505 '510 Wing His Phe Val Thr Leu Asn Gly Ser Thr Val Pro Leu Gly Pro Wing 515 520 525 Ser Asp Thr Gly He Leu Asn Pro Asp Gly Tyr Thr Leu Asn Tyr Asn 530 535 540 Glu Tyr He Val Tyr Asn Pro Asn Gln Val Arg Met Arg Tyr Leu Leu 545 550 555 560 Lys Val Gln Phe Asn Phe Leu Gln Leu Trp * 565 570 (2) INFORMATION FOR SEQ ID NO: 3: (i) CHARACTERISTICS OF THE SEQUENCE: (F) LENGTH: 2265 base pairs (G) TYPE: Nucleic acid (H) TYPE OF HEBRA: Monohebra (I) TOPOLOGY: Linear (il) TYPE OF MOLECULE: cDNA (iii) HYPOTHETICAL: No (iv) ANTIPARALELO: No (vi) ORIGINAL SOURCE: (E) TYPE OF TISSUE: Uterus (ix) ) FEATURE: (A) NAME / KEY : CDS (B) LOCATION: 242..1843 (D) OTHER INFORMATION: / product = "poly ADP Ribose Polymerase" (X) DESCRIPTION OF SEQUENCE: SEQ ID NO: 3: TGGGACTGGT CGCCTGACTC GGCCTGCCCC AGCCTCTGCT TCACCCCACT GGTGGCCAAA 60 TAGCCGATGT CTAATCCCCC ACACAAGCTC ATCCCCGGCC TCTGGGATTG TTGGGAATTC 120 TCTCCCTAAT TCACGCCTGA GGCTCATGGA GAGTTGCTAG ACCTGGGACT GCCCTGGGAG 180 GCGCACACAA CCAGGCCGGG TGGCAGCCAG GACCTCTCCC ATGTCCCTGC TTTTCTTGGC 240 C ATG GCT CCA AAG CCG AAG CCC TGG GTA CAG ACT GAG GGC CCT GAG 286 _ ^ _ ^^^ J ^^ _ * I ^. * * ~ * ~ Yt,. . , _. - ^ ¿~ _-- "-« ^ "•» * «, ^ j.«. ^? ? «, 4 j j & ^ s- ^,.

Met Wing Pro Lys Pro Lys Pro Trp Val Gln Thr Glu Pro Glu 575 580 585 .AAG AAG AAG GGC CGG CAG GCA GGA AGG GAG GAG GAC CCC TTC CGC TCC 334 Lys Lys Lys Gly Arg Gln Wing Gly Arg Glu Glu Asp Pro Phe Arg Ser 590 595 600 ACC GCT GAG GCC GCC CTC AAG GCC ATA CCC GCA GAG AAG CGC ATA ATC CGC 382 Thr Wing Glu Wing Leu Lys Wing Pro Pro Wing Glu Lys Arg He He Arg 605 610 615 GTG GAT CCA ACÁ TGT CCA CTC AGC AGC AAC CCC GGG ACC CAG GTG TAT 430 Val Asp Pro Thr Cys Pro Leu Ser Ser Asn Pro Gly Thr Gln Val Tyr 620 625 630 GAG GAC TAC AAC TGC ACC CTG AAC CAG ACC AAC ATC GAG AAC AAC AAC 478 Glu Asp Tyr Asn Cys Thr Leu Asn Gln Thr Asn He Glu Asn Asn Asn 635 640 645 650 AAC AAG TTC TAC ATC ATC CAG CTG CTC CAA GAC AGC P? C CGC TTC TTC 526 Asn Lys Phe Tyr He He Gln Leu Leu Gln Asp Ser Asn Arg Phe Phe 655 660 665 ACC TGC TGG AAC CGC TGG GGC CGT GTG GGA GAG GTC GGC CAG TCA AAG 574 Thr Cys Trp Asn Arg Trp Gly Arg Val Gly Glu Val GLy Gln Ser Lys 670 675 680 ATC AAC CAC TTC ACA AGG CTA GAA GAT AA GCA G AAG GAC TTT GAG AAG 622 He Asn His Phe Thr Arg Leu Glu Asp Ala Lys Lys Asp Phe Glu Lys 685 690 695 AAA TTT CGG GAA AAG ACC AAG AAC AAC TGG GCA GAG CGG GAC CAC TTT 670 Lys Phe Arg Glu Lys Thr Lys Asn Asn Trp Wing Glu Arg Asp His Phe 700 705 710 GTG TCT CAC CCG GGC AAG TAC ACT CTT ATC GAA GTA CAG GCA GAG GAT 718 Val Ser His Pro Gly Lys Tyr Thr Leu He Glu Val GJn Wing Glu Asp 715 720 725 730 GAG GCC CAG GAA GCT GTG GTG AAG GTG GAC AGA GGC CCA GTG AGG ACT 766 Glu Wing Gln Glu Wing Val Val Lys Val Asp Arg Gly Pío Val Arg Thr 735 740 745 GTG ACT AAG CGG GTG CAG CCC TGC TCC CTG GAC CCA GCC ACG CAG AAG 814 Val Thr Lys Arg Val Gln Pro Cys Ser "Leu Asp Pro Wing Thr Gln Lys 750 755 760 CTC ATC ACT AAC ATC TTC AGC AAG GAG ATG TTC AAG AAC ACC ATG GCC 862 Leu He Thr Asn He Phe Ser Lys Glu Met Phe Lys A = n Thr Met Wing 765 770 775 CTC ATG GAC CTG GAT GTG AAG AAG ATG CCC CTG GGA AAG CTG AGC AAG 910 Leu Met Asp Leu Asp Val Lys Lys Met Pro Leu Gly Lys Leu Ser Lys 780 785 790 CAA CAG ATT GCA CGG GGT TTC GAG GCC GG TTG GAG GCG CTG GAG GCC GCC 958 Gln Gln He Wing Arg Gly Phe Glu Wing Leu Glu Wing Leu Glu Glu Wing 795 800 805. 810 CTG AAA GGC CCC ACG GAT GGT GGC CAG AGC CTG GAG GAG CTG TCC TCA 1006 Leu Lys Gly Pro Thr Asp Gly Gly Gln Ser Leu Glu Glu Glu Leu Ser Ser • 815 820 825 CAC TTT TAC ACC GTC ATC CCG CAC AAC TTC GGC CAC AGC CAG CCC CCG 1054 His Phe Tyr Thr Val He Pro His Asn Phe Gly His Ser Gln Pro Pro 830 835 890 CCC ATC AAT TCC CCT GAG CTT CTG CAG GCC AAG AAG GAC ATG CTG CTG 1102 Pro He Asn Ser Pro Glu Leu Leu Gln Ala Lys Lys Asp Met Leu Leu 845 850 855 GTG CTG GCG GAC ATC GAG CTG GCC CAG GCC CTG CAG GCA GTC TCT GAG 1150 Val Leu Wing Asp He Glu Leu Wing Gln Wing Ala Leu Gln Wing Val Ser Glu 860 865 • 870 CAG GAG AAG ACG GTG GAG QAG GTG CCA CAC CCC CTG GAC CGA CAC TAC 1198 Gln Glu Lys Thr Val Glu Glu Val Pro His Pro Leu Asp Arg Asp Tyr 875 880 885 890 CAG CTT CTC AAG TGC CAG CTG CAG CTG CTA GAC TCT GGA GCA CCT GAG 1246 Gln Leu Leu Lys Cys Gln Leu Gln Leu Leu Asp Ser Gly Ala Pro Glu 895 900 905 TAC AAG GTG ATA CAG ACC TAC TTA GAA CAG ACT GGC AGC AAC CAC AGG 1294 Tyr Lys Val He Gln Thr Tyr Leu Glu Gln Thr Gly Ser Asn His Arg 910 915 920 TGC CCT ACÁ CTT CAÁ CAC ATC TGG AAA GTA AAC CAA GAA GGG GAG GAA 1342 Cys Pro Thr Leu Gln His He Trp Lys Val Asn Gln Glu Glu Glu 925 930 935 GAC AGA TTC CAG GCC CAC TCC AAA CTG GGT AAT CGG AAG CTG CTG TGG 1390 Asp Arg Phe Gln Ala His Ser Lys Leu Gly Asn Arg Lys Leu Leu Trp 940 945 950 CAT GGC ACC AAC ATG GCC GTG GTG GCC GCC ATC CTC ACT AGT GGG CTC 1438 His Gly Thr Asn Met Wing Val Val Wing Wing He Leu Thr 5er Gly Leu 955 960 965 970 CGC ATC ATG CCA CAT TCT GGT GGG CGT GTT GGC AAG GGC ATC TAC TTT 1486 Arg He Met Pro His Ser Gly Gly Arg Val Gly Lys Gly He Tyr Phe 975 980 985 GCC TCA GAG AAC AGC AAG TCA GCT GGA TAT GTT ATT GGC ATG AAG TGT 1534 Wing Ser Glu Asn Ser Lys Ser Wing Gly Tyr Val He Gly Met Lys Cys 990 995 1000 GGG GCC CAC CAT GTC GGC TAC ATG TTC CTG GGT GAG GTG GCC CTG GGC 1582 Gly Wing His His Val Gly Tyr Met Phe Leu Gly Glu Val Wing Leu Gly 1005 1010 1015 AGA GAG CAC CAT ATC AAC ACG GAC AAC CCC AGC TTG AAG AGC CCA CCT 1630 Arg Glu His His As Asn Thr Asp Asn Pro Ser Leu Lys Ser Pro Pro 1020 1025 1030 CCT GGC TTC GAC AGT GTC ATT GCC CGA GGC CAC ACC GAG CCT GAT CCG * 1678 Pro Gly Phe Asp Ser Val He Wing Arg Gly His Thr Glu Pro Asp Pro 1035 1040 1045 1050 f AtJí * -. ^^^^^ ACC CAG GAC ACT GAG TTG GAG CTG GAT GGC CAG CAG GTG GTG GTG CCC 1726 Thr Gln Asp Thr Glu Leu Glu Leu Asp Gly Gln Gln Val Val Val Pro 1055 1060 1065 CAG GGC CAG CCT GTG CCC TGC CCA GAG TTC AGC AGC TCC AC T TCC TCC 1774 Gln Gly Gln Pro Val Pro Cys Pro Glu Phe Ser Ser Ser Thr Phe Ser i 1070 1075 1080 CAG AGC GAC TAC CTC ATC TAC CAG GAG AGC CAG TGT CGC CTG CGC TAC 1822 Gln Ser Glu Tyr Leu He Tyr Gln Glu Ser Gln Cys Arg Leu Arg Tyr 1085 1090 1095 CTG CTG GAG GTC CAC CTC TGA GTGCCCGCCC TGTCCCCCGG GGTCCTGCAA 1873 Leu Leu Glu Val His Leu * 1100 1105 GGCTGGACTG TGATCTTCAA TCATCCTGCC CATCTCTGGT ACCCCTATAT CACTCCTTTT 1933 TTTCAAGAAT ACAATACGTT GTTGTTAACT ATAGTCACCA TGCTGTACAA GATCCCTGAA 1993 CTTATGCCTC CTAACTGAAA TTTTGTATTC TTTGACACAT CTGCCCAGTC CCTCTCCTCC 2053 CAGCCCATGG TAACCAGCAT TTGACTCTTT ACTTGTATAA GGGCAGCTTT TATAGGTTCC 2113 ACATGTAAGT GAGATCATGC AGTGTTTGTC TTTCTGTGCC TGGCTTATTT CACTCAGCAT 2173 AATGTGCACC GGGTTCACCC ATGTTTTCAT AAATGACAAG ATTTCCTCCT TTAAAAAAAA 2233 AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AA 2265 (2) INFORMATION FOR SEQ ID NO: 4: (1) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 534 amino acids (B) TYPE: amino acid (D) TOPOLOGY: Linear (n) TYPE OF MOLECULE: Protein (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO. 4: Met Wing Pro Lys Pro Lys Pro Trp Val Gln Thr Glu Pro Glu Lys 1 5 10 15 Lys Lys Gly Arg Gln Wing Gly Arg Glu "Glu Asp Pro Phe Arg Ser Thr 20 25 30 Ala Glu Ala Leu Lys Ala He Pro Wing Glu Lys Arg He He Arg Val 35 40 45 Asp Pro Thr Cys Pro Leu Ser As Asn Pro Gly Thr Gln Val Tyr Glu 50 55 60 Asp Tyr Asn Cys Thr Leu Asn Gln Thr Asn He Glu Asn Asn Asn Asn 65 70 75 80 Lys Phe Tyr He He Gln Leu Leu Gln Asp Ser Asn Arg Phe Phe Thr 85 90 95 Cys Trp Asn Arg Trp Gly Arg Val Gly Val Val Gly Gla Ser Lys He j & g Jte ^ 1 1 100 105 110 Asn His Phe Thr Arg Leu Glu Asp Ala Lys Lys Asp Phe Glu Lys Lys 115 120 125 Phe Arg Glu Lys Thr Lys Asn Asn Trp Wing Glu Arg Asp His Phe Val 130 135 140 Ser His Pro Gly Lys Tyr Thr Leu He Glu Val Gln Wing Glu Asp Glu 145 150 155 160 Wing Gln Glu Wing Val Val Lys Val Asp Arg Gly Pro Val Arg Thr Val 165 190 175 Thr Lys Arg Val Gln Pro Cys Ser Leu Asp Pro Wing Thr Gln Lys Leu 180 185 190 He Thr Asn He Phe Ser Lys Glu Met Phe Lys Asn Thr Met Ala Leu 195 200 205 Met Asp Leu Asp Val Lys Lys Met Pro Leu Gly Lys Leu Ser Lys Gln 210 215 220 Gln He Wing Arg Gly Phe Glu Wing Leu Glu Wing Leu Glu Glu Wing Leu 225 230 235 240 Lys Gly Pro Thr Asp Gly Gly Gln Ser Leu Glu Glu Leu Ser Ser His 245 250 255 Phe Tyr Thr Val He Pro His Asn Phe Gly His Ser Gln Pro Pro Pro 260 265 270 He Asn Ser Pro Glu Leu Glu Ala Lys Lys Asp Met Leu Leu Val 275 280 285 Leu Wing Asp He Glu Leu Wing Gln Wing Leu Gln Wing Val Ser Glu Gln 290 295 300 Glu Lys Thr Val Glu Glu Val Pro His Pro Leu Asp Arg Asp Tyr Gln 305 310 315 320 Leu Leu Lys Cys Gln Leu Gln Leu Leu Asp Ser Gly Wing Pro Glu Tyr 325 330 335 Lys Val He Gln Thr Tyr Leu Glu Gln Thr Gly Ser Asn His Arg Cys 340 345 350 Pro Thr Leu Gln His He Trp Lys Val Asn Gln Glu Glu Glu Glu Asp 355 360 365 Arg Phe Gln Wing His Ser Lys Leu Gly Asn Arg Lys Leu Leu Trp His 370 375 380 Gly Thr Asn Met Wing Val Val Wing Wing He Leu Thr Ser Gly Leu Arg 385 390 395 40Ó He Met Pro His Ser Gly Gly Arg Val Gly Lys Gly He Tyr Phe Wing 405 910 415 BÜ MMMÜ Ser Glu Asn Ser Lys Ser Wing Gly Tyr Val He Gly Met Lys Cys Gly 420 425 430 Wing His His Val Gly Tyr Met Phe Leu Gly Glu Val Wing Leu Gly Arg 435 440 445 Glu His His As Asn Thr Asp As Ser Leu Lys Pro Pro Pro 450 455 460 Gly Phe Asp Ser Val He Wing Arg Gly His Thr Glu Ero Asp Pro Thr 465 470 475 480 Gln Asp Thr Glu Leu Glu Leu Asp Gly Gln Gln Val Val Val Pro Gln 485 490 495 Gly Gln Pro Val Pro Cys Pro Glu Phe Ser Ser Ser Thr Phe Ser G-ln 500 505 510 Ser Glu Tyr Leu He Tyr Gln Glu Ser Gln Cys Arg Leu Arg Tyr Leu 515 520 525 Leu Glu Val His Leu * 10 530 (2) INFORMATION FOR SEQ ID NO: 5: (1) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 2265 base pairs (B) TYPE: Nucleic acid (C) TYPE OF HEBRA: Monohebra (D) TOPOLOGY: Linear (n) TYPE OF MOLECULE: cDNA 15 (V) HYPOTHETIC: No (vi) ANTIPARALELO: No (vi) ORIGINAL SOURCE: (F) TYPE OF TISSUE: Uterus (x) CHARACTERISTIC: (A) NAME / KEY: CDS (B) LOCATION: 221. .1843 '(D) OTHER INFORMATION ÓN: / product = "pol? ADP Ribose Polymerase "20 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 5: TGGGACTGGT CGCCTGACTC GGCCTGCCCC AGCCTCTGCT TCACCCCACT GGTGGCCAAA 60 TAGCCGATGT CTAATCCCCC ACACAAGCTC ATCCCCGGCC TCTGGGATTG TTGGGAATTC 120 TCTCCCTAAT TCACGCCTGA GGCTCATGGA GAGTTGCTAG ACCTGGGACT GCCCTGGGAG 180 GCGCACACAA CCAGGCCGGG TGGCAGCCAG GACCTCTCCC ATG TCC CTG CTT TTC 235 Met Ser Leu Leu Phe 535 25 TTG GCC ATG GCT CCA AAG CCG AAG CCC TGG GTA CAG ACT GAG GGC CCT 283 ^ g ^^^^^^^^^ | ^ ga ^^^^^ a ^^^^^^ - ^ s4 ^^^^^^ | ^^ i? i ^ i? i ^ _ ^^ ^^^ á ¿íai Leu Ala Met Wing Pro Lys Pro Lys Pro Trp Val Gln Thr Glu Gly Pro 540 545 550 555 GAG AAG AAG AAG GGC CGG CAG GCA GGA AGG GAG GAG GAC CCC TTC CGC 331 Glu Lys Lys Gly Gly Lg Wing Gly Arg Glu Glu Asp Pro Phe Arg 560 565 570 TCC ACC GCT GAG GCC CTC AAG GCC ATA CCC GCA GAG AAG CGC ATA ATC 379 Ser Thr Ala Glu Ala Leu Lys Ala He Pro Ala Glu Lys Arg He He 575 580 585 CGC GTG GAT CCA ACA TGT CCA CTC AGC AGC AAC CCC GGG ACC CAG GTG 427 Arg Val Asp Pro Thr Cys Pro Leu 5er Ser Asn Pro Gly Thr Gln Val 590 - 595 600 TAT GAG GAC TAC AAC TGC ACC CTG AAC CAG ACC AAC ATC GAG AAC AAC 475 Tyr Glu Asp Tyr Asn Cys Thr Leu Asn Gln Thr Asn He Glu Asn Asn 605 610 615 AAC AAC AAG TTC TAC ATC ATC CAG CTG CTC CAA GAC AGC AAC CGC TTC 523 Asn Asn Lys Phe Tyr He He Gln Leu Leu Gln Asp Ser Asn Arg Phe 620 625 630 635 TTC ACC TGC TGG AAC CGC TGG GGC CGT GTG GGA GAG GTC GGC CAG TCA 571 Phe Thr Cys Trp Asn Arg Trp Gly Arg Val Gly Glu Va l Gly Gln Ser 640 645 650 AAG ATC AAC CAC TTC ACA AGG CTA GAA GAT GAC AAG AAG GAC TTT GAG 619 Lys He Asn His Phe Thr Arg Leu Glu Asp Ala Lys Lys Asp Phe Glu 655 660 665 AAG AAA TTT CGG GAA AAG ACC AAG AAC AAC TGG GCA GAG CGG GAC CAC 667 Lys Lys Phe Arg Glu Lys Thr Lys Asn Asn Trp Wing Glu Arg Asp His 670 675 680 TTT GTG TCT CAC CCG GGC AAG TAC AT CTT ATC GAA GTA CAG GCA GAG 715 Phe Val Ser His Pro Gly Lys Tyr Thr Leu He Glu Val Gln Ala Glu 685 690 695 GAT GAG GCC CAG GAA GCT GTG GTG AAG GTG GAC AGA GGC CCA GTG AGG 763 Asp Glu Ala Gln Glu Ala Val Val Lys Vai Asp Arg Gly Pro Val Arg 700 705 710 715 ACT GTG ACT AAG CGG GTG CAG CCC TGC TCC CTG GAC CCA GCC ACG CAG 811 Thr Val Thr Lys Arg Val Gln Pro Cys Ser Leu Asp Pro Ala Thr Gln 720 725 '730 AAG CTC ATC ACT AAC ATC TTC AGC AAG GAG ATG TTC AAG AAC ACC ATG 859 Lys Leu He Thr Asn He Phe Ser Lys Glu Met Phe Lys Asn Thr Met 735 740 '745 GCC CTC ATG GAC CTG GAT GTG AAG AAG ATG CCC CTG GGA AAG CTG AGC 907 Wing Leu Met Asp Leu Asp Val Lys Lys Met Pro Leu Gly Lys Leu Ser- 750 755 760 AAG CAG ATT GCA CGG GGT TTC GAG GCC TTG GAG GCG CTG GAG GAG 955 Lys Gln Gln He Ala Arg Gly Phe Glu Ala Leu Glu Ala Leu Glu Glu 765 770 775 • & *. ,. .. **, ..¿ .... «,. , ..,,. ....., ._ ,, »... , ._ .. ".1A« 1a «att ?. tt aMito GCC CTG AAA GGC CCC ACG GAT GGT GGC CAA AGC CTG GAG GAG CTG TCC 1003 Wing Leu Lys Gly Pro Thr Asp Gly Gly Gln Ser Leu G..u Glu Leu Ser 780 785 790 795 TCA CAC TTT TAC ACC GTC ATC CCG CAC AAC TTC GGC CAC AGC CAG CCC 1051 Ser His Phe Tyr Thr Val He Pro His Asn Phe Gly H_s Ser Gln Pro 800 805 810 CCG CCC ATC AAT TCC CCT GAG CTT CTG CAG GCC AAG AAG GAC ATG CTG 1099 Pro Pro He Asn Ser Pro Glu Leu Leu Gln Wing Lys Lys Asp Met Leu 815 820 825 CTG GTG CTG GCG GAC ATC GAG CTG GCC CAG GCC CTG CAG GCA GTC TCT 1147 Leu Val Leu Wing Asp He Glu Leu Wing Gln Wing Leu Gln Wing Val Ser 830 835 840 GAG CAG GAG AAG ACG GTG GAG GAG GTG CCA CAC CCC CTG GAC CGA GAC 1195 Glu Gln Glu Lys Thr Val Glu Glu Val Pro His Pro Leu Asp Arg Asp 845 850 855 TAC CAG CTT CTC AAG TGC CAG CTG CAG CTG CTA GAC TCT GGA GCA CCT 1243 Tyr Gln Leu Leu Lys Cys Gln Leu Gln Leu Leu Asp Ser Gly Wing Pro 860 865 870 875 GAG TAC AAG GTG ATA CAG ACC TAC TTA GAA CAG ACT GGC AGC AAC CAC 1291 Glu Tyr Lys Val He Gln Thr Tyr Leu Glu Gln Thr GLy Ser Asn His 880 885 890 AGG TGC CCT ACÁ CTT CAÁ CAC ATC TGG AAA GTA AAC CAA GAA GGG GAG 1339 Arg Cys Pro Thr Leu Gln His He Trp Lys Val Asn GLn Glu Gly Glu 895 900 905 GAA GAC AGA TTC CAG GCC CAC TCC AAA CTG GGT AAT CGG AAG CTG CTG 1387 Glu Asp Arg Phe Gln Wing H s Ser Lys Leu Gly Asn Acg Lys Leu Leu 910 915 920 TGG CAT GGC ACC AAC ATG GCC GTG GTG GCC GCC ATC CTC ACT AGT GGG 1435 Trp His Gly Thr Asn Met Wing Val Val Wing Wing He Lsu Thr Ser Gly 925 930 935 CTC CGC ATC ATG CCA CAT TCT GGT GGG CGT GTT GGC A °? G GGC ATC TAC 1483 Leu Arg He Met Pro His Ser Gly Gly Arg Val Gly Lys Gly He Tyr 940 945 950 955 TTT GCC TCA GAG AAC AGC AAG TCA GCT GGA TAT GTT ATT GGC ATG AAG 1531 Phe Wing Ser Glu Asn Ser Lys Ser Wing Gly Tyr Val He Gly Met Lys 960 965 970 TGT GGG GCC CAC CAT GTC GGC TAC ATG TTC CTG GGT G «VG GTG GCC CTG 1579 Cys Giy Wing His His Val Gly Tyr Met Phe Leu Gly Glu Val Wing Leu 975 980 985 GGC AGA GAG CAC CAT ATC AAC ACG GAC AAC CCC AGC TTG AAG AGC CCA 1627 Gly Arg Glu His His He Asn Thr Asp Asn Pro Ser Leu Lys Ser Pro 990 995 1000 CCT CCT GGC TTC GAC AGT GTC ATT GCC CGA GGC CAC ACC GAG CCT GAT 1675 Pro Pro Gly Phe Asp Ser Val He Wing Arg Gly His Thr Glu Pro Asp , í * í. Í.? HÍ, '.- AÍ: J¿? R.Át.J > :, i, ^ _, ^? g ^ ái ^ L mA 1005 1010 1015 CCG ACC CAG GAC ACT GAG TTG GAG CTG GAJf 'GGC CAG CAG GTG GTG GTG 1723 Pro Thr Gln Asp Thr Glu Leu Glu Leu Asp Gly Gln Gln Val Val Val 1020 1025 1030 1035 CCC CAG GGC CAG CCT GTG CCC TGC CCA GAG TTC AGC AGC TCC ACA TTC 1771 Pro Gln Gly Gln Pro Val Pro Cys Pro Glu Phe Ser Ser Ser Thr Phe 1040 1095 1050 5 TCC CAG AGC GAG TAC CTC ATC TAC CAG GAG AGC CAG TGT CGC CTG CGC 1819 Ser Gln Ser Glu Tyr Leu He Tyr Gln Glu Ser Gln Cys Arg Leu Arg 1055 1060- 1065 TAC CTG GAG GTC CAC .CTC TGA GTGCCCGCCC TGTCCCCCGG GGTCCTGCAA 1873 Tyr Leu Leu Glu Val His Leu * 1070 1075 GGCTGGBCTG TGATCTTCAA TCATCCTGCC CATCTCTGGT ACCCCTATAT CACTCCTTTT 1933 TTTCAAGAAT ACAATACGTT GTTGTTAACT ATAGTCACCA TGCTGTACAA GATCCCTGAA 1993 CTTATGCCTC CTAACTGAAA TTTTGTATTC TTTGACACAT CTGCCCAGTC CCTCTCCTCC 2053 CAGCCCATGG TAACCAGCAT TTGACTCTTT ACTTGTATAA GGGCAGCTTT TATAGGTTCC 2113 ACATGTAAGT GAGATCATGC AGTGTTTGTC TTTCTGTGCC TGGCTTATTT CACTCAGCAT 2173 AATGTGCACC GGGTTCACCC ATGTTTTCAT AAATGACAAG ATTTCCTCCT TTAAAAAAAA 2233 AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AA 2265 (2) INFORMATION FOR SEQ ID NO: 6: 15 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 541 amino acids (B) TYPE: amino acid (D) TOPOLOGY: Linear (ii) TYPE OF MOLECULE: Protein (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 6: Met Ser Leu Leu Phe Leu Ala Met Pro Wing Lys Pro Lys Pro Trp Val 1 5 10 15 Gln Thr Glu Pro Glu Lys Lys Lys Gly Arg Gln Wing Gly Arg Glu 20 20 25 . 30 Glu Asp Pro Phe Arg Ser Thr Ala Glu Ala Leu Lys Ala He Pro Ala 35 40 45 Glu Lys Arg He He Arg Val Asp Pro Thr Cys Pro Leu Ser As As 50 55 60 Pro Gly Thr Gln Val Tyr Glu Asp Tyr Asn Cys Thr Leu Asn Gln Th'r 65 70 75 80 Asn He Glu Asn Asn Asn Asn Lys Phe Tyr He He Gln Leu Leu Gln 5 85 90 95 "" - • - '"• - • ** Asp Ser Asn Arg Phe Phe Thr Cys Trp Asn Arg Trp Gly Arg Val Gly 100 105 110 Glu Val Gly Gln Ser Lys He Asn His Phe Thr Arg Leu Glu Asp Ala 115 120 125 Lys Lys Asp Phe Glu Lys Lys Phe Arg Glu Lys Thr Lys Asn Asn Trp 130 135 140 Wing Glu Arg Asp His Phe Val Ser His Pro Gly Lys Tyr Thr Leu He 145 '150 155 160 Glu Val Gln Wing Glu Asp Glu Wing Gln Glu Wing Val Val Lys Val Asp 165 170 175 Arg Gly Pro Val Arg Thr Val Thr Lys Arg Val Gln Pro Cys Ser Leu 180 185 190 Asp Pro Wing Thr Gln Lys Leu He Thr Asn He Phe Ser Lys Glu Met 195 200 205 Phe Lys Asn Thr Met Ala Leu Met Asp Leu Asp Val Lys Lys Met Pro 210 215 220 Leu Gly Lys Leu Ser Lys Gln Gln He Ala Arg Gly Phe Glu Ala Leu 225 230 235 240 Glu Ala Leu Glu Glu Ala Leu Lys Gly Pro Thr Asp Gly Gly Gln Ser 245 250 255 Leu Glu Glu Leu Ser Ser His Phe Tyr Thr Val He Pro His Asn Phe 260 265 270 Gly His Ser Gln Pro Pro Pro He Asn Ser Pro Glu Leu Glu Ala 275 280 285 Lys Lys Asp Met Leu Leu Val Leu Ala Asp He Glu Leu Wing Gln Wing 290 295 300 Leu Gln Wing Val Ser Glu Gln Glu Lys Thr Val Glu Glu Val Pro His 305 310 315 320 Pro Leu Asp Arg Asp Tyr Gln Leu Leu "Lys Cys Gln Leu Gln Leu Leu 325 330 335 Asp Ser Gly Wing Pro Glu Tyr Lys Val He Gln Thr Tyr Leu Glu Gln 340 345 350 Thr Gly Ser Asn His Arg Cys Pro Thr Leu Gln His He Trp Lys Val 355 360 365 Asn Gln Glu Gly Glu Glu Asp Arg Phe Gln Wing His Ser Lys Leu Gly 370 375 380 Asn Arg Lys Leu Leu Trp His Gly Thr Asn Met Wing Val Val Wing Wing 385 390 395 400 He Leu Thr Ser Gly Leu Arg He Met Pro His Ser Gly Gly Arg Val 405 910 415 ^ _ ^^^^^^^^^^^^ * ^^^ faith «^? ^^^^^ rfj * ^^^» áy * ^^ g ^^^^ * Astg ^ g ^^ _ ^ ____ ^ «j¡ ^^^^^ __ ^ .___ ^^ ^^^^^^ Gly Lys Gly íle Tyr Phe Wing Ser Glu Asn Ser Lys 5er Wing Gly Tyr 420 425 í:" 430 Val He Gly Met Lys Cys Gly Ala His His Val Gly Tyr Met Phe Leu 435 440 445 Gly Glu Val Ala Leu Gly Arg Glu His His As Asn Thr Asp Asn Pro 450 455 460 Ser Leu Lys Pro Pro Pro Gly Phe Asp Ser Val He Ala Arg Gly 5 465 470 975 480 His Thr Glu Pro Asp Pro Thr Gln Asp Thr Glu Leu Glu Leu Asp Gly 485 •. 490 495 Gln Gln Val Val Val Pro Gln Gly Gln Pro Val Pro Cys Pro Glu Phe 500 505 510 Ser Ser Thr Phe Ser Gln Ser Glu Tyr Leu He Tyr Gln Glu Ser 515 520 525 0 Gln Cys Arg Leu Arg Tyr Leu Leu Glu Val His Leu * 530 535 540 (2) INFORMATION FOR SEQ ID NO: 7: (i) CHARACTERISTICS OF THE SEQUENCE : (A) LENGTH: 1740 base pairs (B) TYPE: Nucleic acid (C) TYPE OF HEBRA: Monohebra (D) TOPOLOGY: Linear (li) TYPE OF MOLECULE: cDNA 15 (lil) HYPOTHETIC: No (iv) ANTIPARALELO : I did not see) ORIGINAL SOURCE: (A) ORGANISM: Mus musculus (ix) FEATURE: (A) NAME / KEY: CDS (B) LOCATION: 112..1710 20 (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 7: CCCGGCTTTC ACTTTTTCTG CTGCCTCGGG GAACACCTCG AGCCAACTGC TTCCTAACTC AGGGTGGGCA GAACTGACGG GATCTAAGCT TCTGCATCTC TGAGGAGAAC C ATG GCT 117 Met Wing CCA AAA CGA AAG GCC TCT GTG CAG ACT GAG GGC TCC AAG AAG CAG CGA 165 Pro Lys Arg Lys Ala Ser Val Gln Thr Glu Gly Ser Lys Lys Gln Arg 545 550 555 25 1 CAA GGG ACÁ GAG GAG GAG GAC AGC TTC CGG TCC ACT GCC GAG GCT GCT CTC 213 Gln Gly Thr Glu Glu Glu Asp Ser Phe Arg Ser Thr Ala Glu Ala Leu 560 565 570 575 AGA GCA GCC CCT GCT GAT AAT CGG GTC ATC CGT GTG GAC CCC TCA TGT 261 Arg Wing Wing Pro Wing Asp Asn Arg Val He Arg Val Asp Pro Ser Cys 580 585 590 CCA TTC AGC CGG AAC CCC GGG ATA CAG GTC CAC GAG GAC TAT GAC TGT 309 Pro Phe Ser Arg A = n Pro Gly He Gln Val His Glu Asp Tyr Asp Cys 5 595 600 605 ACC CTG AAC CAG ACC AAC ATC GGC AAC AAC AAC AAC AAG TTC TAT ATT 357 Thr Leu Asn Gln Thr Asn He Gly Asn Asn Asn Lys Phe Tyr He 610 615 620 ATC CAÁ CTG CTG GAG GAG GGT AGT CGC TTC TTC TGC TGG AAT CGC TGG 405 He Gln Leu Leu Glu Glu Gly Ser Arg Phe Phe Cys Tip Asn Arg Trp 625 630 635 GGC CGG GTG GGA GAG GTG GGC CAG AGC AAG ATG AAC CAC TTC ACC TGC 453 Gly Arg Val Gly Glu Val Gly Gln Ser Lys Met Asn His Phe Thr Cys 10 640 645 650 655 CTG GAA GAT GAG AAG AAG GAC TTT AAG AAG AAA TTT TGG GAG AAG ACT 501 Leu Glu Asp Ala Lys Lys Asp Phe Lys Lys Lys Phe Trp Glu Lys Thr 660 665 670 AAA AAC AAA TGG GAG GAG CGG GAC CGT TTG GTG GCC CAG CCC AAC AAG 549 Lys Asn Lys Trp Glu Glu Arg Asp Arg Phe Val Wing Gln Pro Asn Lys 675 680 685 TAC ACA CTT ATA GAA GTC CAG GGA GAA GCA GAG AGC CAA GAG GCT GTA 597 Tyr Thr Leu He Glu Val Gln Gly Glu Wing Glu Ser Gln Glu Wing Val 15 690 695 700 GTG AAG GCC TTA TCT CCC CAG GTG GAC AGC GGC CCT GTG AGG ACC GTG 645 Val Lys Ala Leu Ser Pro Gln Val Asp Ser Gly Pro Val Arg Thr Val 705 710 715 GTC AAG CCC TGC TCC CTA GAC CCT GCC ACC CAG AAC CTT ATC ACC AAC 693 Val Lys Pro Cys Ser Leu Asp Pro Ala Thr Gln Asn Leu He Thr Asn 720 725 - 730 735 ATC TTC AGC AAA GAG ATG TTC AAG AAC GCA ATG ACC CTC ATG AAC CTG 741 He Phe Ser Lys Glu Met Phe Lys Asn Ala Met Thr Leu Met Asn Leu 20 740 745 750 GAT GTG AAG AAG ATG CCC TTG GGA AAG CTG ACC AAG CAG CAG ATT GCC 789 Asp Val Lys Lys Met Pro Leu Gly Lys Leu Thr Lys Gln Gln He Wing 755 760 765 CGT GGC TTC GAG GCC TTG GAA T CTA GAG GAG GCC ATG AAA AAC CCC 837 Arg Gly Phe Glu Ala Leu Glu Ala Leu Glu Glu Ala Met Lys Asn Pro 770 775 780 ACA GGG GAT GGC CAG AGC CTG GAA GAG CTC TCC TCC TGC TTC TAC ACT Thr Gly Asp Gly Gln Ser Leu Glu Glu Leu Ser Ser Cys Phe Tyr Thr 25 785 790 795 GTC ATC CCA CAC AAC TTC GGC CGC AGC CGA CCC CCG CCC ATC AAC TCC 933 Val He Pro His Asn Phe Gly Arg Ser Arg Pro Pro Pro He Asn Ser 800 805 810 815 CCT GAT GTG CTT CAG GCC AAG AAG GAC ATG CTG CTG GTG CTA GCG GAC 981 Pro Asp Val Leu Gln Wing Lys Lys Asp Met Leu Leu Val Leu Ma Asp 820 825 830 5 ATC GAG TTG GCG CAG ACC TTG CAG GCC GCC CCT GGG GAG GAG GAG GAG 1029 He Glu Leu Wing Gln Thr Leu Gln Wing Pro Pro Gly Glu Glu Glu Glu 835 840 * 845 AAA GTG GAA GAG GTG CCA CAC CCA CTG GAT CGA GAC TAC CAG CTC CTC 1077 Lys Val Glu Val Val Pro His Pro Leu Asp Arg Asp Tyr Gln Leu Leu 850 855 860 AGG TGC CAG CTT CAA CTG CTG GAC TCC GGG GAG TCC GAG TAC AAG GCA 1125 Arg Cys Gln Leu Gln Leu Leu Asp Ser Gly Glu Ser Glu Tyr Lys Wing 865 870 875 10 ATA CAG ACC TAC CTG AAA CAG ACT GGC AAC AGC TAC AGG TGC CCA AAC 1173 He Gln Thr Tyr Leu Lys Gln Thr Gly Asn Ser Tyr Arg Cys Pro Asn 880 885 890 895 CTG CGG CAT GTT TGG AAA GTG AAC CGA GAA GGG GAG GGA GAC AGG TTC 1221 Leu Arg His Val Trp Lys Val Asn Arg Glu Gly Glu Gly Asp Arg Phe 900 905 910 CAG GCC CAC TCC AAA CTG GGC AAT CGG AGG CTG CTG TGG CAC GGC ACC 1269 Gln Ala His Ser Lys Leu Gly Asn Arg Arg Leu Lep Trp His Gly Thr 915 920 925 15 AAT GTG GCC GTG GTG GCT GCC ATC CTC ACC AGT GGG CTC CGA ATC ATG 1317 Asn Val Ala Val Val Ala Ala He Leu Thr Ser Gly Leu Arg He Met 930 935 940 CCA CAC TCG GGT GGT CGT GTT GGC AAG GGT ATT TAT TTT GCC TCT GAG 1365 Pro His Ser Gly Gly Arg Val Gly Lys Gly He Tyr Phe Wing Ser Glu 945 950 955 AAC AGC AAG TCA GCT GGC TAT GTT ACC ACC ATG CAC TGT GGG GGC CAC 1413 Asn Ser Lys Ser Wing Gly Tyr Val Thr Thr Met His Cys Gly Gly His 960 965 970 975 20 CAG GTG GGC TAC ATG TTC CTG GGC GAG GTG GCC CTC GGC AAA GAG CAC 1461 Gln Val Gly Tyr Met Phe Leu Gly Glu Val Ala Leu Gly Lys Glu His 980 985 990 CAC ATC ACC ATC GAT GAC CCC AGC TTG AAG AGT CCA CCC CCT GGC TTT 1509 His He Thr He Asp Asp Pro Ser Leu Lys Ser Pro Pro Pro Gly Phe 995 1000 1005 GAC AGC GTC ATC GCC CGA GGC CA GAG CCG GAT CC C GCC CAG GAC 1557 Asp Ser Val He Wing Arg Gly Gln Thr Glu Pro Asp Pro Wing Gln Asp 1010 1015 1020 2 ATT GAA CTT GAA CTG GAT GGG CAG CCG GTG GTG GTG CCC CAA GGC CCG 1605 Tímu ^ SMáí &a ^ te ^^ He Glu Leu Glu Leu Asp Gly Gln Pro Val Val Val Pro Gln Gly Pro 1025 1030 1035 CCT GTG CAG TGC CCG TCA TTC AAA AGC TCC AGC TTC AGC CAG AGT GAA 1653 Pro Val Gln Cys Pro Ser Phe Lys Ser Ser Ser Phe Ser Gln Ser Glu 1040 1045 1050 1055 TAC CTC ATA TAC AAG GAG AGC CAG TGT CGC CTG CTG CTG CTG GAG 1701 Tyr Leu He Tyr Lys Glu Ser Gln Cys Arg Leu Arg Tyr Leu Leu Glu 1060 1065 1070 5 ATT CAC CTC TAAGCTGCTT GCCCTCCCTA GGTCCAAGCC 1740 He His Leu (2) INFORMATION FOR SEQ ID NO: 8: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 533 amino acids (B) TYPE: amino acid (D) TOPOLOGY: Linear (ii) TYPE OF MOLECULE: protein 10 (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 8: Met Ala Pro Lys Arg Lys Ma Ser Val Gln Thr Glu Gly Ser Lys Lys 1 5 10 15 Gln Arg Gln Gly Thr Glu Glu Glu Asp Ser Phe Arg Ser Thr Ma Glu 20 25 30 Wing Leu Arg Ma Ma Pro Ma Asp Asn Arg Val He Arg Val Asp Pro 35 40 45 Ser Cys Pro Phe Ser Arq Asn Pro Gly He Gln Val His Glu Asp Tyr 15 50 55 60 Asp Cys Thr Leu Asn Gln Thr Asn He Gly Asn Asn Asn Asn Lys Phe 65 70 75 80 Tyr He He Gln Leu Leu Glu Glu Gly Be Arg Phe Phe Cys Trp Asn 85 90 95 Arg Trp Gly Arg Val Gly Glu Val Gly-. Gln Ser Lys Met Asn His Phe 100 105 110 Thr Cys Leu Glu Asp Ma Lys Lys Asp Phe Lys Lys Lys Phe Trp Glu 20 115 120 125 Lys Thr Lys Asn Lys Trp Glu Glu Arg Asp Arg Phe Val Ala Gln Pro 130 135 140 Asn Lys Tyr Thr Leu He Glu Val Gln Gly Glu Ma Glu Ser Gln Glu 145 150 155 160 Ma Val Val Lys Ala Leu Ser Pro Gln Val Asp Ser Gly Pro Val Arg 165 170 175 Thr Val Val Lys Pro Cys Ser Leu Asp Pro Ma Thr Gln Asn Leu He 25 180 185 190 Thr Asn He Phe Ser Lys Glu Met Phe Lys Asn Ma Met Thr Leu Met 195 200 205 Asn Leu Asp Val Lys Lys Met Pro Leu Gly Lys Leu Thr Lys Gln Gln 210 215 220 He Ala Arg Gly Phe Glu Ma Leu Glu Wing Leu Glu Glu Wing Met Lys 225 230 235 240 Asn Pro Thr Gly Asp Gly Gln Ser Leu Glu Glu Leu Ser Ser Cys Phe 245 2-50 255 Tyr Thr Val He Pro His Asn Phe Gly Arg Ser Arg Pro Pro Pro He 260 265 270 Asn Ser Pro Asp Val Leu Gln Ma Lys Lys Asp Met Leu Leu Val Leu 275 280 285 Wing Asp He Glu Leu Ma Gln Thr Leu Gln Wing Ma Pro Gly Glu Glu 290 295 300 Glu Glu Lys Val Glu Glu Val Pro His Pro Leu Asp Arg Asp Tyr Gln 305 310 315 320 Leu Leu Arg Cys Gln Leu Gln Leu Leu Asp Ser Gly Glu Ser Glu Tyr 325 330 335 Lys Ma He Gln Thr Tyr Leu Lys Gln Thr Gly Asn Ser Tyr Arg Cys 340 345 350 Pro Asn Leu Arg His Val Trp Lys Val Asn Arg Glu Gly Glu Gly Asp 355 360 365 Arg Phe Gln Wing His Ser Lys Leu Gly Asn Arg Arg Leu Leu Trp His 370 375 380 Gly Thr Asn Val Ma Val Val Ma Ala He Leu Thr Ser Gly ^ eu Arg 385 390 395 400 He Met Pro His Ser Gly Gly Arg Val Gly Lys Gly He Tyr? He Wing 405 410 415 Ser Glu Asn Ser Lys Ser Wing Gly Tyr Val Thr Thr Met His Cys Gly 420 425 430 Gly His Gln Go Gly Tyr Met Phe Leu Gly Glu Val Ma Leu Gly Lys 435 440 445 Glu His His He Thr He Asp Asp Pro Ser Leu Lys Ser Pro Pro Pro 450 455 460 Gly Phe Asp Ser Val He Ma Arg Gly Gln Thr Glu Pro Asp Pro Ma 465 470 475 480 Gln Asp He Glu Leu Glu Leu Asp Gly Gln Pro Val Val Val Pro Gln 485 490 495 Gly Pro Pro Val Gln Cys Pro Ser Phe Lys Ser Ser Ser Phe Ser Gln 500 505 510 Ser Glu Tyr Leu He Tyr Lys Glu Ser Gln Cys Arg Leu Arg Tyr Leu 515 520 525 Leu Glu He His Leu 530 • (2) INFORMATION SEQ ID NO: 9: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 1587 base pairs (B) TYPE: Nucleic acid (C) TYPE OF HEBRA: Monohebra (D) TOPOLOGY: Linear (ii) TYPE OF MOLECULE : cDNA (iii) HYPOTHETICAL: No (iv) ANTIPARALELO: No (vi) ORIGINAL SOURCE: (A) ORGANISM: Mus musculus (ix) CHARACTERISTIC: (B) NAME / KEY: CDS (B) LOCATION: 1..1584 ( xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 9: ATG GCT CCA AAA CGA AAG GCC TCG GTG CAG ACT GAG GGC TCC AAG AAG 48 Met Wing Pro Lys Arg Lys Ma Ser Val Gln Thr Glu Gly Ser Lys Lys 535 540 545 CAG CGA CAGA GGG ACÁ GAG GAG GAG GAC AGC TTC CGG TCC ACT GCC GAG 96 Gln Arg Gln Gly Thr Glu Glu Glu Asp Ser Phe Arg Ser Thr Ala Glu 550 555 560 565 GCT CTC AGA GCA GCC CCT GCT GAT AAT CGG GTC ATC CGT GTG GAC CCC 144 Ma Leu Arg Ma Ma Pro Ma Asp Asi, Arg Val He Arg Val Asp Pro 570 575 580 TCA TGT CCA TTC AGC CG AAC CCC GGG ATA CAG GTC CAC GAG GAC TAT 192 Ser Cys Pro Phe Ser Arg Asn Pro Gly He Gln Val His Glu Asp Tyr 585 590 595 GAC TGT ACC CTG AAC CAG ACC AAC ATC GGC AAC AAC AAC AAC AAG TTC 240 Asp Cys Thr Leu Asn Gln Thr Asn He Gly Asn Asn Asn Asn Lys Phe 600 605 610 TAT ATT ATC CA CT CTG GAG GAG GGT AGT CGC TTC TTC TGC TGG AAT 288 Tyr ile He Gln Leu Leu Glu Glu Gly Be Arg Phe Phe Cys Trp Asn 615 620 625 CGC TGG GGC CGC GTG GGA GAG GTG GGC CAG AGC AAG ATG AAC CAC TTC 336 Arg Trp Gly Arg Val Gly Glu Val Gly Gln Ser Lys Met Asn His Phe 630 635 640 645 ACC TGC CTG GAA GAT GCA AAG AAG GAC TTT AAG AAA AAA TTT TGG GAG 384 Thr Cys Leu Glu Asp Ma Lys Lys Asp Phe Lys Lys Lys Phe Trp Glu 650 655 660 AAG ACT AAA AAC AAA TGG GAG GAG CGG GAC CGT TTG GTG GCC CAG CCC 432 Lys Thr Lys Asn Lys Trp Glu Glu Arg Asp Arg Phe Val Ma Gin Pro 665 670 675 AAC AAG TAC ACA CTT ATA GAA GTC CAG GGA GAA GCA GAG AGC CAG GAG 480 Asn Lys Tyr Thr Leu He Glu Val Gln Gly Glu Glu Glu Ser Gln Glu 680 - 685 690 GCT GTA GTG AAG GTG GAC AGC GGC CCT GTG AGG ACC GTG GTC AAG CCC 528 Wing Val Val Lys Val Asp Ser Gly Pro Val Arg Thr Val Val Lys Pro 695 700 705 TGC TCC CTA GAC CCT GCC ACC CAG AAC CTT ATC ACC AAC ATC TTC AGC 576 Cys Ser Leu Asp Pro Ma Thr Gln Asn Leu He Thr Asn He Phe Ser 710 715 720 725 AAA GAG ATG TTC AAG AAC GCA ATG ACC CTC ATG AAC CTG GAT GTG AAG 624 Lys Glu Met Phe Lys Asn Wing Met Thr Leu Met Asn Leu A sp Val Lys 730 735 740 AAG ATG CCC TTG GGA AAG CTG ACC AAG CAG CAG ATT GCC CGT GGC TTC 672 Lys Met Pro Leu Gly Lys Leu Thr Lys Gln Gln He Ma Arg Gly Phe 745 750 755 GAG GCC TTG GAA GCT CTA GAG GAG GCC ATG AAA AAC CCC ACA GGG GAT 720 Glu Ala Leu Glu Ma Leu Glu Glu Ma Met Lys Asn Pro Thr Gly Asp 760 765 770 GGC CAG AGC CTG GAA GAG CTC TCC TCC TGC TTC TAC ACT GTC ATC CCA 768 Gly Gln Ser Leu Glu Glu Leu Ser Ser Cys Phe Tyr Thr Val He Pro 775 780 785 CAC AAC TTC GGC CGC AGC CGA CCC CCG CCC ATC AAC TCC CCT GAT GTG 816 His Asn Phe Gly Arg Be Arg Pro Pro Pro He Asn Ser Pro Asp Val 790 795 800 805 CTT CAG GCC AAG AAG GAC ATG CTG CTG GTG CTA GCG GAC ATC GAG TTG 864 Leu Gln Ma Lys Lys Asp Met Leu Leu Val Leu Wing Asp He Glu Leu 810 815 • 820 GCG CAG ACC TTG CAG GCC GCC CCT GGG GAG GAG GAG GAG AAA GTG GAA 912 Wing Gln Thr Leu Gln Ma Ma Pro Gly Glu Glu Glu Glu Lys Val Glu 825 830 • 835 GAG GTG CCA CAC CCA CTG GAT CGA GAC TAC CAG CTC CTC AGG TGC CAG 960 Glu Val Pro His Pro Leu Asp Arg Asp Tyr Gln Leu Leu Arg Cys Gln 840 845 850 CTT CA CT CT G GCC TCC GGG GAG TCC GAG TAC AAG GCA ATA CAG ACC 1008 Leu Gln Leu Leu Asp Ser Gly Glu Ser Glu Tyr Lys Ma He Gln Thr 855 860 865 ^^^^ - ** "*" * • '*' «a» * »- a * - TAC CTG AAA CAG ACT GGC AAC AGC TAC AGG TGC CCA A ^ C CTG CGG CAT 1056 Tyr Leu Lys Gln Thr Gly Asn Ser Tyr Arg Cys Pro Asn Leu Arg His 870 875 880 885 GTT TGG AAA GTG AAC CGA GAA GGG GAG GAC AGC TGC CAG GCC CAC 1104 Val Trp Lys Val Asn Arg Glu Gly Glu Gly Asp Arg Phe Gln Ma His 890 895 900 TCC AAA CTG GGC AAT CGG AGG CTG CTG TGG CAC GGC ACC AAT GTG GCC 1152 Ser Lys Leu Giy Asn Arg Arg Leu Leu Trp H s Gly Thr Asn Val Wing 905 910 915 GTG GTG GCT GCC ATC CTC ACC AGT GGG CTC CGA ATC AGG CCA CAC TCG 1200 Val Val Ma Ma He Leu Thr Ser Gly Leu Arg He M = t Pro His Ser 920 925 930 GGT GGT CGT GTT GGC AAG GGT ATT TAT TTT GCC TCT GAG AAC AGC AAG 1248 Gly Gly Arg Val Gly Lys Gly He Tyr Phe Ma Ser Glu Asn Ser Lys 935 940 945 TCA GCT GGC TAT GTT ACC ACC ATG CAC TGT GGG GGC CAC CAG GTG GGC 1296 Ser Ma Gly Tyr Val Thr Thr Met His Cys Gly Gly HLS Gln Val Gly 950 955 960 965 TAC ATG TTC CTG GGC GAG GTG GCC CTC GGC AAA GAG CAC CAC ATC ACC 1344 Tyr Met Phe Leu Gly Val Wing Ala Leu Gly Lys Glu HLS H s He Thr 970 975 980 ATC GAT GAC CCC AGC TTG AAG AGT CCA CCC CCT GGC TTT GAC AGC GTC 1392 He Asp Asp Pro 5er Leu Lys Ser Pro Pro Pro Gly Phe Asp Ser Val 985 990 995 ATC GCC CGA GGC CAG ACC GAG CCG GAT CCC GCC CAG GAC ATT GAA CTT 1490 He Wing Arg Gly Gln Thr Glu Pro Asp Pro Wing Gln Asp He Glu Leu 1000 1005 101C) GAA CTG GAT GGG CAG CCG GTG GTG GT CA CCC CAG GGC CCG CCT GTG CAG 148 I Glu Leu Asp Gly Gln Pro Val Val Val Pro Gln Gly Pro Pro Val Gln 1015 1020 1025 TGC CCG TCA TTC AAA AGC TCC AGC TTC AGC CAG AGT GAA TAC CTC ATA 1536 Cys Pro Ser Phe Lys Ser Ser Ser Phe Ser Gln Ser Giu Tyr Leu He 1030 1035 109C 1 1045 TAC AAG GAG AGC CAG TGT CGC CTG CGC TAC CTG CTG GAG ATT CAC CTC 1584 Tyr Lys Glu Ser Gln Cys Arg Leu Arg Tyr Leu Leu G.Lu He His Leu 1050 1055 1060 TAA 1587 (2) INFORMATION FOR SEQ ID NO: 10: (1) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 528 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (n) TYPE OF MOLECULE: Protein 25 (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 10: ^^ ^ ^ ^ ^ ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Wing Ser Val Gln? Hr Glu Gly Ser Lys Lys 1 5 10 15 Gln Arg Gln Gly Thr Glu Glu Glu Asp Ser Phe Arg Ser Thr Ma Glu 20 25 30 Ma Leu Arg Ala Ala Pro Ma Asp Asn Arg Val He Arg Val Asp Pro 35 40 45 Ser Cys Pro Phe Ser Arg Asn Pro Gly He Gln Val His Glu Asp Tyr 50 55 60 Asp Cys Thr Leu Asn Gln Thr Asn He Gly Asn Asn Asn Asn Lys Phe 65 70, 75 80 Tyr He He Gln Leu Leu Glu Glu Gly Ser Arg Phe Phe Cys Trp Asn 85 90 95 Arg Trp Gly Arg Val Gly Glu Val Gly Gln Ser Lys Met Asn His Phe 100 105 110 10 Thr Cys Leu Glu Asp Ma Lys Lys Asp Phe Lys Lys Lys Phe Trp Glu 115 120 125 Lys Thr Lys Asn Lys Trp Glu Glu Arg Asp Arg Phe Val Ma Gln Pro 130 135 140 Asn Lys Tyr Thr Leu He Glu Val Gln Giy Glu Wing Glu Ser Gln Glu 145 150 155 160 Wing Val Val Lys Val Asp Ser Gly Pro Val Arg Thr Val Val Lys Pro 165 170 175 15 Cys Ser Leu Asp Pro Wing Thr Gln Asn Leu He Thr Asn He Phe Ser 18 0 185 190 Lys Glu Met Phe Lys Asn Ma Met Thr Leu Met Asn Leu Asp Val Lys 195 200 205 Lys Met Pro Leu Gly Lys Leu Thr Lys Gln Gln He Ma Arg Gly Phe 210 215 220 Glu Ma Leu Glu Ma Leu Glu Glu Ma Met Lys Asn Pro Thr Gly Asp 225 230 235 240 Gly Gln Ser Leu Glu Glu Leu Be Ser Cys Phe Tyr Thr Val He Pro 245 250 255 His Asn Phe Gly Arg Ser Arg Pro Pro Pro He Asn Ser Pro Asp Val 260 265. 270 Leu Gin Ma Lys Lys Asp Met Leu Leu Val Leu Wing Asp He Glu Leu 275 280 285 Wing Gln Thr Leu Gln Ala Wing Pro Gly Glu Glu Glu Glu Lys Val Glu 290 295 300 25 Glu Val Pro His Pro Leu Asp Arg Asp Tyr Gln Leu Leu Arg Cys Gln 305 310 315 320 Leu Gln Leu Leu Asp Ser Gly Glu Ser Glu Tyr Lys Ma He Gln Thr 325 330 335 Tyr Leu Lys Gln Thr Gly Asn Ser Tyr Arg Cys Pro Asn Leu Arg His 340 345 350 Val Trp Lys Val Asn Arg Glu Gly Glu Gly Asp Arg Phe Gln Ala His 355 360 365 Ser Lys Leu Gly Asn Arg Arg Leu Leu Trp His Gly Thr Asn Val Ma 370 375 380 Val Val Ala Ma He Leu Thr Ser Giy Leu Arg He Met Pro His Ser 385 390 395 400 Gly Gly Arg Val Gly Lys Gly He Tyr Phe Ma Ser Glu Asn Ser Lys 405 410 415 Ser Ma Gly Tyr Val Thr Thr Met His Cys Gly Gly HLS Gln Val Gly 420 425 430 10 Tyr Met Phe Leu Gly Glu Val Ma Leu Gly Lys Glu HLS His He Thr 435 440 445 He Asp Asp Pro Ser Leu Lys Ser Pro Pro Pro Gly Phe Asp Ser Val 450 455 460 He Ma Arg Gly Gln Thr Glu Pro Asp Pro Ma Gln Asp He Glu Leu 465 470 475 480 Glu Leu Asp Gly Gln Pro Val Val Val Pro Gln Pro Pro Val Gln 485 490 495 15 Cys Pro Ser Phe Lys Ser Ser Phe Ser Gln Ser Glu Tyr Leu He 500 505 510 Tyr Lys Glu Ser Gln Cys Arg Leu Arg Tyr Leu Leu Glu He His Leu 515 520 525 (2) PAR INFORMATION? SEQ ID NO: 11: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 14 amino acids (B) TYPE: amino acid 20 (C) TYPE OF HEBRA: Monohebra (D) TOPOLOGY: Linear (ii) TYPE OF MOLECULE: peptide (lil) HYPOTHETIC: Yes (IX) CHARACTERISTICS: (A) NAME / KEY: Region (B) LOCATION: 2 (C) OTHER INFORMATION: / note = "Xaa steht fuer 1 bis 5 andere ~ t- Aminosaeuren" (ix) FEATURE: (A) NAME / KEY: Region (B) LOCATION: 3 (C) OTHER INFORMATION: / note = "Xaa steht fuer Ser oder Thr" (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 11: Pro Xaa Xaa Gly Xaa Xaa Xaa Gly Lys Gly He Tyr Phe Wing 1 5 10 (2) INFORMATION FOR SEQ ID NO: 12: (I) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 amino acids (B) TYPE: amino acid ( C) TYPE OF HEBRA: Monohebra (D) TOPOLOGY: Linear (II) TYPE OF MOLECULE: peptide (m) HYPOTHETIC: Yes 10 (x) CHARACTERISTICS: (A) NAME / KEY: Region (B) LOCATION: 1 (D) OTHER INFORMATION: / pota = "Xaa steht fuer Ser oder Thr" (IX) CHARACTERISTICS: (A) NAME / KEY: Region (B) LOCATION: 6 (C) OTHER INFORMATION: / note = "Xaa steht fuer He oder Val" (ix) CHARACTERISTICS: 15 (A) NAME / KEY: Region (B) LOCATION: 9 (D) OTHER INFORMATION: / note = "Xaa steht fuer 1 bis 5 andere Ammosaeuren" (ix) FEATURE: (A) NAME / KEY: Region (B) LOCATION: 10 (D) OTHER INFORMATION: / note = "Xaa steht fuer Ser oder Thr" (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 12: 20 Xaa Xaa Gly Leu Arg Xaa Xaa Pro Xaa Xaa Gly Xaa Xaa Xaa Gly Lys 1 5 10 - 15 Gly He Tyr Phe Ma 20 (2) INFORMATION FOR SEQ ID NO: 13: (1) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 45 amino acids 25 ^ (B) TYPE: amino acid (C) TI PO OF HEBRA: Mcf fS #? Ebra (D) TOPOLOGY: Linear * (ii) TYPE OF MOLECULE: peptide (iii) HYPOTHETICAL: Si (ix) "CHARACTERISTIC: (A) NAME / KEY: Region (B) LOCATION: 16 (D) OTHER INFORMATION: / note = "Xaa steht fuer Ser oder Thr" (ix) FEATURE: (A) NAME / KEY: Region (B) LOCATION: 21 (D) OTHER INFORMATION: / note = "Xaa steht fuer He oder Val" (ix) CHARACTERISTICS: (A) NAME / KEY: Region (B) LOCATION: 24 (D) OTHER INFORMATION: / note = "Xaa steht fuer 1a 5 andere 10 A inosaeuren" (ix) FEATURE: (A) NAME / KEY: Region (B) LOCATION: 25 (D) OTHER INFORMATION: / note = "Xaa steht fuer Ser oder Thr" (ix) CHARACTERISTICS: (A) NAME / KEY: Region (B) LOCATION: 6 (D) OTHER INFORMATION : / note = "Xaa steht fuer Ser oder Thr" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 1 13: Leu Leu Trp His Gly Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa He Leu Xaa 1 5 10 15 Xaa Gly Leu Arg Xaa Xaa Pro Xaa Xaa Gly Xaa Xaa Xaa Gly Lys Gly 20 25 30 He Tyr Phe Ala Xaa Xaa Ser Lys '' Ser Ma Xaa Tyr 35 40 45 20 (2) INFORMATION FOR SEQ ID NO: 13: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 22 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: Monoblend (D) ) TOPOLOGY: Linear (li) TYPE OF MOLECULE: peptide (iii) HYPOTHETIC: Yes 25 (ix) FEATURE: (A) "NAME / KEY: Region (B) LOCATION: 1 (D) OTHER INFORMATION: / note =" Xaa steht fuer Leu oder Val "(xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 14 : Xaa Xaa Xaa Xaa Xaa Xa Xau Leu Xaa Xaa Xaa Xaa Xaa Xaa Leu Xaa 1 5 10 L5 Xaa Xaa Xaa Xaa Xaa Leu 20 (2) INFORMATION FOR SEQ -ID NO: 15: (i) CHARACTERISTICS OF THE SEQUENCE: (A) ) LENGTH: 27 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: Monohebra (D) TOPOLOGY: Linear (ii) TYPE OF MOLECULE: peptide 0 (iii) HYPOTHETICAL: Si (ix) CHARACTERISTIC: (A) NAME / KEY: Region (B) LOCATION: 21 (E) OTHER INFORMATION: / note = "Xaa steht fuer Asp oder Glu" (ix) CHARACTERISTICS: (C) NAME / KEY: Region (D) LOCATION: 22 (D) OTHER INFORMATION : / note = "Xaa steht fuer 10 oder 11 andere 5 Aminosaeuren" (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 15: Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Tyr Xaa Xaa 1 5 10 15 Gln Leu Leu Xaa Xaa Xaa Trp Gly Arg Val Gly 20 25 (2) INFORMATION FOR SEQ ID NO: 15: 0 (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 29 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: Monohebra (D) TOPOLOGY: Linear (ii) TYPE OF MOLECULE: peptide (iii) HYPOTHETIC: Yes ^^ ^ j ^ ggg ^ • i í «SS (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 16: "Wing Xaa Xaa Xaa Phe Xaa Lys Xaa Xaa Xaa Xaa Lys Tbr Xaa Asn Xaa 1 5 10 15 Trp Xaa Xaa Xaa Xaa Xaa Phe Xaa Xaa Xaa Pro Xaa Lys 20 25 (2) INFORMATION FOR SEQ ID NO: 17: (l) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 44 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: Monohebra (D) TOPOLOGY: Linear (i) TYPE OF MOLECULE: peptide (iii) HYPOTHETIC: Yes (ix) CHARACTERISTICS: (A) NAME / KEY: Region (B) LOCATION: 4 (D) OTHER INFORMATION: / note = "Xaa steht fuer He oder Leu" (Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 17: Gln Xaa Leu Xaa Xaa Xaa He Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa 1 5 10 15 Met Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Pro Leu Gly Lys Leu 20 25 30 Xaa Xaa Xaa Gln He Xaa Xaa Xaa Xaa Xaa Xaa Leu 35 40 (2) INFORMATION FOR SEQ ID NO: 18: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 15 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: Monohebra (D) TOPOLOGY: Linear ( ii) TYPE OF MOLECULE: peptide (iii) HYPOTHETIC: Yes (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 18: Phe Tyr Thr Xaa He Pro His Xaa Phe Gly Xaa Xaa Xaa Pro Pro 1 5 10 15 (2) INFORMATION FOR SEQ ID MO: 19: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 17 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: Monohebra (D) TOPOLOGY: Linear (n) TYPE OF MOLECULE: peptide (m) HYPOTHETICAL: Si ( xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 19: Lys Xaa Xaa Xaa Leu Xaa Xaa Leu Xaa Asp He Glu Xaa Ala Xaa Xaa 1 5 10 15 (2) INFORMATION FOR SEQ ID NO: 20: (1) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 11 amino acids (B) TYPE : amino acid (C) TYPE OF HEBRA: Monohebra 10 (D) TOPOLOGY: Linear (n) TYPE OF MOLECULE: peptide (lll) HYPOTHETIC: Yes (i) DESCRIPTION OF SEQUENCE: SEQ ID NO: 20: Gly Xaa Xaa Xaa Leu Xaa Glu Val Ma Leu Gly 1 5 10 (2) INFORMATION FOR SEQ ID NO: 21: 15 (l) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: Monohebra (D) TOPOLOGY: Linear (n) TYPE OF MOLECULE: peptide (m) HYPOTHETIC: Si (ix) CHARACTERISTICS: 20 (A) NAME / KEY: Region (B) LOCATION: 14 (D) OTHER INFORMATION: / note = "Xaa steht fuer 7 bis 9 andere Aminosaeuren" (Xl) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 21: Gly Xaa Xaa Ser Xaa Xaa Xaa Xaa Gly Xaa Xaa Xaa Pro Xaa Leu Xaa 1 5 10 15 Gly Xaa Xaa Val 25 ifirf irti ^ MT "# ('1 -fr -f ff frfr y, * tit ^? _? _ ... t ^ t I, aX¿ ^, ^ 20 (2) INFORMATION FOR SEQ ID NO: 22: ( i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 16 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: Monohebra (D) TOPOLOGY: Linear (n) TYPE OF MOLECULE: peptide (ili) HYPOTHETIC: Yes (ix) ) CHARACTERISTICS: (A) NAME / KEY: Region (B) LOCATION: 2 (D) OTHER INFORMATION: / note = "Xaa steht fuer Tyr oder Phe" (l) SEQUENCE DESCRIPTION: SEQ ID NO: 22: Glu Xaa Xaa Xaa Tyr Xaa Xaa Xaa Gln Xaa Xaa Xaa Xaa Tyr Leu Leu 1 5 10 15 (2) INFORMATION FOR SEQ ID NO: 23: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 amino acids (B) TYPE : amino acid (C) TYPE OF HEBRA: Monohebra (D) TOPOLOGY: Linear (li) TYPE OF MOLECULE: peptide (ni) HYPOTHETIC: No (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 23: Met Ma Ma Arg Arg Arg Arg Ser Thr Gly Gly Gly Arg Ma Arg Wing 1 5 10 15 Leu Asn Glu Ser 20 (2) INFORMATION FOR SEQ. ID NO: 24: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 20 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: Monohebra (D) TOPOLOGY: Linear (n) TYPE OF MOLECULE: peptide (nor ) HYPOTHETICAL: No (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 24: Lys Thr Glu Leu Gln Ser Pro Glu His Pro Leu Asp Gln His Tyr Arg 1 5 10 15 Asn Leu His Cys 20 (2) INFORMATION FOR SEQ ID NO: 25: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 21 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: Monohebra (D) TOPOLOGY: Linear (ii) TYPE OF MOLECULE: peptide (iii) HYPOTHETIC: No (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 25: Cys Lys Gly Arg Gln Wing Gly Arg Glu Glu Asp Pro Phe Arg Ser Thr 1 5 10 15 Wing Glu Ma Leu Lys 20 (2) INFORMATION FOR SEQ ID NO: 26: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 20 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: Monohebra (D) TOPOLOGY: Linear (ii) TYPE OF MOLECULE: peptide (iii) HYPOTHETIC: No (xi) DESCRIPTION OF SEQUENCE: SEQ ID NO: 26: Cys Lys Gln Gln He Ma Arg Gly Phe Glu Ma Leu Glu Ma Leu Glu 1 5 10 LS Glu Ma Leu Lys 20 (2) INFORMATION FOR SEQ ID NO: 27: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: Monohebra (D) TOPOLOGY: Linear (ii) TYPE OF MOLECULE: peptide * * ¿- * (iii) HYPOTHETICAL: No (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 27: Lys Gln Gln He Ma Arg Gly Phe Glu Ma Leu Glu M.a Leu Glu Glu 1 '5 10 15 Ala Leu Lys (2) INFORMATION FOR SEQ ID NO: 28: (i) CHARACTERISTICS OF THE SEQUENCE: (A) LENGTH: 19 amino acids (B) TYPE: amino acid (C) TYPE OF HEBRA: Monoblend (D) TOPOLOGY: Linear (ii) TYPE OF MOLECULE: peptide (iii) HYPOTHETIC: No (xi) DESCRIPTION OF THE SEQUENCE: SEQ ID NO: 28: Lys Gln Gln He Ma Arg Gly Phe Glu Ma Leu Glu Ala Leu Glu Glu 1 5 10 15 Ala Met Lys

Claims (9)

1. A poly (ADP-ribose) polymerase (PARP) homologue from a human or non-human mammal, which has 5 an amino acid sequence with: a) a functional NAD + binding domain and b) a motif of the finger sequence other than zinc, of the general formula: CX2CXmHX2C in which m is an integer value from 28 or 30, and the radicals X are, independent of each other, any Ciminoacid.

2. The PARP homolog as claimed in claim 15, wherein the NAD + binding domain comprises one of the following general sequence motifs: PXn (S / T) GX3GKGIYFA, (S / T) XGRL (I / V) XPXn (S / T) GX3GKGIYFA or LL HG (S / T) X7IL (S / T) XGLR (I / V) XPXn (S / T) GX3GKGIYFAX3SKSAXY 20 in which n is an integer value from 1 to 5, and the X radicals are, independent of each other, any amino acid.

3. The PARP homologue, as claimed in any of the preceding claims, consisting of when 25 minus one of the following sequence reasons partial: LX9NX2YX2Q LX (D / E) X10 / ?? GRVG, AX3FXKX4KTXNX X5FX3PXK, QXL (I / L) X2IX9MX10PLGKLX3QIX6L, FYTXIPHXFGX3PP; and KX3LX2LXDIEXAX2L, in which the X radicals are, independent of each other, any amino acid. . The PARP homolog as claimed in any of the preceding claims, selected from the human PARP homologs, which has the amino acid sequence shown in SEQ ID NO: 2 (human PARP2) or SEQ ID NO: 4 6 6 (human PARP3 type 1 6 2; or murine PARP homologs which have the amino acid sequence shown in SEQ ID NO: 8 (long form of mouse PARP) or SEQ ID NO: 10 (short form of Mouse PARP) A binding counterpart with a specificity for the PARP homologs as claimed in any of the preceding claims, selected from: a) antibodies and fragments thereof, b) protein-like compounds that interact with a partial sequence of the protein, and c) molecular weight effectors under which they modulate the catalytic activity of PARP or another function ^^^ "^ yí ^^ Sii ^^ K ^^ J ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ ^^^^^^^^? ^ T ^^^^^ jj biological of a PARP molecule. 6. A nucleic acid containing: a) a nucleotide sequence encoding at least one PARP homologue, as claimed in any one of claims 1 to 4, or the nucleotide sequence complementary thereto; b) a nucleotide sequence that hybridizes to a sequence as specified in a) under stringent conditions; or c) the nucleotide sequences that come from the nucleotide sequences defined in a) and b) through the degeneracy of the genetic code. 7. The nucleic acid as claimed in claim 6, which contains: a) nucleotides +3 to +1715 shown in SEQ ID NO: 1; b) nucleotides +242 to +1843 shown in SEQ ID NO: 3; c) nucleotides +221 to +1843 shown in SEQ ID NO: 5; d) nucleotides +112 to +1710 shown in SEQ ID NO: 7; or e) nucleotides +1 to +1584 shown in SEQ ID NO: 9. 8. An expression cassette containing, under control J t i i i ¿i i i i _ _ _,, A A A A A A A A A A A A A A A A A A A A A A. »&, .i» .A, ??. ^ ALi > .i ^ a &^ ¡A. < . > M ^^ genetic of at least one nucleotide regulatory sequence at least one nucleotide sequence as claimed in any of claims 6 or 7. 9. A recombinant vector containing at least one expression cassette as claimed in claim 8 10. A recombinant microorganism containing at least one recombinant vector as claimed in claim 9. 11. A transgenic mammal containing a vector as claimed in claim 9. 12. A mammal deficient in PARP or a deficient eukaryotic cell. in PARP, in which the functional expression of at least one gene coding for a PARP homolog is inhibited as claimed in any of claims 1 to

4. 13. An in vitro detection method for PARP inhibitors, the which consists of: a) incubating a target capable of reacting polyADP-ribosylation with support or without support, with a reaction mixture that contains: a) a PARP homolog as claimed in any of claims 1 to 4, a2) a PARP activator; Y a3) a PARP inhibitor or an analyte in which at least one PARP inhibitor is suspected; b) performing the polyADP-ribosylation reaction; and c) determining the polyADP-ribosylation of the target in a qualitative or quantitative manner. The method as claimed in claim 13, wherein the PARP homolog is preincubated with the PARP activator and the PARP inhibitor or an analyte in which at least one PARP inhibitor is suspected, prior to 10 perform the polyADP-ribosylation reaction. 1

5. The method as claimed in any of claims 13 and 14, wherein the target capable of polyADP-ribosylation reaction is a histone protein. 1

6. The method as claimed in any of claims 13 to 15, wherein the PARP activator is activated DNA. 1

7. The method as claimed in any of claims 13 to 16, wherein the reaction of 20 polyADP-ribosylation is initiated by adding NAD +. 1

8. The method as claimed in any of claims 13 to 17, wherein the polyADP-ribosylation of the supported target is determined using anti-poly (ADP-ribose) antibodies. 1

9. The method as claimed in any of the claims 13 to 17, wherein the unsupported target is labeled with a fluorophore acceptor. The method as claimed in claim 19, wherein the polyADP-ribosylation of the unsupported target is determined using an anti-poly (ADP-ribose) antibody labeled with a donor fluorophore capable of transferring energy to the acceptor fluorophore. The method as claimed in any of claims 19 and 20, wherein the target is biotinylated histone, and the acceptor fluorophore is coupled thereto via avidin or streptavidin. 2. The method as claimed in any of claims 20 and 21, wherein the antipole antibody (ADP-ribose) carries a europium crypt as a donor fluorophore. 3. An in vitro detection method for binding counterparts for a molecule of PARP, which consists of: al) immobilizing at least one PARP homologue as claimed in any of claims 1 to 4 on a support; bl) contacting the immobilized PARP homologue with an analyte in which at least one binding counterpart is suspected; and cl) determine, as appropriate after an incubation period, the constituents of the analyte bound to the immobilized PARP homolog; or a2) immobilizing on an support an analyte containing at least one possible binding partner for a molecule of PARP; b2) contacting the immobilized analyte with at least one PARP homologue or claimed in any of claims 1 to 4 for which a binding partner is sought; and c2) examining the immobilized analyte, as appropriate after the incubation period, for the binding of the PARP homolog. 4. A method for the qualitative or quantitative determination of the nucleic acids encoding a PARP homologue as claimed in any of claims 1 to 4, which consists of: a) incubating a biological sample with a defined amount of an acid Exogenous nucleic acid as claimed in any of claims 6 and 7, hybridizing under stringent conditions, determining the hybridizing nucleic acids, as appropriate, comparing with a standard; or b) incubating a biological sample with a pair of oligonucleotide primers with specificity for a nucleic acid encoding a homologue of ^^^^ ^ ^^^^^^^^^^^^^^^^^^^^ ^ ^ ^ ^^^^^^^^^^^^^^^^^^ ^ g ^ ?? ^^^^^^ b ^^ j ^ kJjA ^^ Í PARP, amplify the nucleic acid, determine the product of the amplification and, as appropriate, compare with a standard. 25. A method for the qualitative or quantitative detection of a PARP homologue as claimed in any of claims 1 to 4, which consists of: a) incubating a biological sample with a specific binding partner for a PARP homologue, b) detect the binding partner / PARP complex and, as appropriate, c) compare the result with a standard. 26. The method as claimed in claim 25, wherein the binding counterpart is an antibody or a binding fragment thereof, which bears a detectable label, as appropriate. 27. The method as claimed in any of claims 24 to 26 for diagnosing diseases mediated by energy deficiency. 28. A method for determining the efficacy of PARP effectors, which consists in: a) incubating a PARP homologue as claimed in any of claims 1 to 4 with an analyte consisting of an effector of a physiological or pathological condition; Separate the effector again, as appropriate; Y b) determine the activity of the PARP homolog, as appropriate, after adding the substrates or co-substrates. 29. A composition for gene treatment containing an acceptable vehicle for gene treatment, a nucleic acid construct, which: a) contains an antisense nucleic acid against a coding nucleic acid as claimed in any of claims 6 and 7; or b) a ribozyme against a nucleic acid as claimed in any of claims 6 and 7; or c) codes for a specific PARP inhibitor. 30. A pharmaceutical composition containing, in a carrier acceptable for pharmaceutical use, at least one PARP protein as claimed in any of claims 1 to 4, at least one binding partner of PARP as claimed in claim 5 or when minus a nucleotide coding sequence as claimed in claim 6 or 7. 31. The use of the low molecular weight PARP binding counterparts as claimed in claim 5 for the manufacture of a pharmaceutical agent for diagnosis or treatment of pathological states in the development and / or progress of which is involved when ¡^! ^ ... ^ fe! (W ^ | * gj? E ^ | ^^ teií ^^^ minus one PARP protein, or a polypeptide from it.) 32. The use of PARP binding partners. of low molecular weight, as claimed in claim 5 for the manufacture of a pharmaceutical agent for the diagnosis or treatment of pathological conditions mediated by energy deficiency. ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ ^^^^^^^^ t ^^^ j ^^^^^? ^^