[go: up one dir, main page]

WO1999046294A1 - Gene humain de type chd-1 - Google Patents

Gene humain de type chd-1 Download PDF

Info

Publication number
WO1999046294A1
WO1999046294A1 PCT/CN1998/000036 CN9800036W WO9946294A1 WO 1999046294 A1 WO1999046294 A1 WO 1999046294A1 CN 9800036 W CN9800036 W CN 9800036W WO 9946294 A1 WO9946294 A1 WO 9946294A1
Authority
WO
WIPO (PCT)
Prior art keywords
polypeptide
cbfbjb02
seq
polynucleotide
nucleotide sequence
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CN1998/000036
Other languages
English (en)
Inventor
Mao Mao
Juan Zhou
Jisheng Wu
Qinghua Zhang
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shanghai Second Medical University
Original Assignee
Shanghai Second Medical University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shanghai Second Medical University filed Critical Shanghai Second Medical University
Priority to PCT/CN1998/000036 priority Critical patent/WO1999046294A1/fr
Publication of WO1999046294A1 publication Critical patent/WO1999046294A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals

Definitions

  • This invention relates to newly identified polynucleotides, polypeptides encoded by them and to the use of such polynucleotides and polypeptides, and to their production. More particularly, the polynucleotides and polypeptides of the present invention relate to the DNA binding protein family, hereinafter referred to as CBFBJB02. The invention also relates to inhibiting or activating the action of such polynucleotides and polypeptides.
  • the human global transcription activator homologous sequence and the mouse chromodomain- helicase-DNA binding protein (CHD-1) are members of the DNA binding protein family and are thought to play a role in gene regulation. This indicates that the DNA binding protein family has an established, proven history as therapeutic targets. Clearly there is a need for identification and characterization of further members of the DNA binding protein family which can play a role in preventing, ameliorating or correcting dysfunctions or diseases, including, but not limited to, cancer, autoimmune disease, and AIDS.
  • the invention relates to CBFBJB02 polypeptides and recombinant materials and methods for their production. .Another aspect of the invention relates to methods for using such CBFBJB02 polypeptides and polynucleotides. Such uses include the treatment of cancer, autoimmune disease, and AIDS, among others.
  • the invention relates to methods to identify agonists and antagonists using the materials provided by the invention, and treating conditions associated with CBFBJB02 imbalance with the identified compounds.
  • diagnostic assays for detecting diseases associated with inappropriate CBFBJB02 activity or levels are examples of diseases associated with inappropriate CBFBJB02 activity or levels.
  • CBFBJB02 refers, among others, generally to a polypeptide having the amino acid sequence set forth in SEQ ID NO:2 or an allelic variant thereof.
  • CBFBJB02 activity or CBFBJB02 polypeptide activity or “biological activity of the CBFBJB02 or CBFBJB02 polypeptide” refers to the metabolic or physiologic function of said CBFBJB02 including similar activities or improved activities or these activities with decreased undesirable side-effects Also included are antigenic and lmmunogenic activities of said CBFBJB02
  • CBFBJB02 gene refers to a polynucleotide having the nucleotide sequence set forth m SEQ ID NO 1 or allehc variants thereof and/or their complements
  • Antibodies as used herein includes polyclonal and monoclonal antibodies, chime ⁇ c, single chain, and humanized antibodies, as well as Fab fragments, mcluding the products of an Fab or other lmmunoglobuhn expression library
  • Isolated means altered “by the hand of man” from the natural state If an "isolated” composition or substance occurs in nature, it has been changed or removed from its original environment, or both For example, a polynucleotide or a polypeptide naturally present m a living animal is not “isolated ' but the same polynucleotide or polypeptide separated from the coexisting mate ⁇ als of its natural state is "isolated", as the term is employed herein
  • Polynucleotide generally refers to any poly ⁇ bonucleotide or polydeox ⁇ bonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA
  • Polynucleotides include, without limitation single- and double-stranded DNA, DNA that is a mixture of single- and double- stranded regions, single- and double-stranded RNA, and RNA that is mixture of single- and double-stranded regions, hybrid molecules comprising DNA and RNA that may be single-stranded or, more typically, double-stranded or a mixture of single- and double-stranded regions
  • polynucleotide refers to t ⁇ ple-stranded regions comprising RNA or DNA or both RNA and DNA
  • the term polynucleotide also includes DNAs or RNAs containmg one or more modified bases and DNAs or RNAs with backbones modified for stability or for other reasons "Modified" bases include, for example, t ⁇
  • a substituted or inserted amino acid residue may or may not be one encoded by the genetic code.
  • a variant of a polynucleotide or polypeptide may be a naturally occurring such as an allelic variant, or it may be a variant that is not known to occur naturally. Non-naturally occurring variants of polynucleotides and polypeptides may be made by mutagenesis techniques or by direct synthesis.
  • Identity is a relationship between two or more polypeptide sequences or two or more polynucleotide sequences, as determined by comparing the sequences.
  • identity also means the degree of sequence relatedness between polypeptide or polynucleotide sequences, as the case may be, as determined by the match between strings of such sequences.
  • Identity and similarity can be readily calculated by known methods, including but not limited to those described in (Computational Molecular Biology, Lesk, A.M. , ed.
  • Preferred methods to determine identity are designed to give the largest match between the sequences tested. Methods to determine identity and similarity are codified in publicly available computer programs. Preferred computer program methods to determine identity and similarity between two sequences include, but are not limited to, the GCG program package (Devereux, J. , et al. , Nucleic Acids Research 12(1): 387 (1984)), BLASTP, BLASTN, and FASTA (Atschul, S.F. et al., /. Molec. Biol. 215: 403-410 (1990). The BLAST X program is publicly available from NCBI and other sources (BLAST Manual, Altschul, S.
  • Preferred parameters for polypeptide sequence comparison include the following: 1) Algorithm: Needleman and Wunsch, J. Mol Biol. 48: 443-453 (1970) Comparison matrix: BLOSSUM62 from Hentikoff and Hentikoff, Proc. Natl. Acad. Sci. USA. 89: 10915-10919 (1992) Gap Penalty: 12 Gap Length Penalty: 4 A program useful with these parameters is publicly available as the "gap" program from Genetics Computer Group, Madison WI. The aforementioned parameters are the default parameters for polypeptide comparisons (along with no penalty for end gaps).
  • Preferred parameters for polynucleotide comparison include the following:
  • Preferred polynucleotide embodiments further include an isolated polynucleotide comprising a polynucleotide having at least a 50,60, 70, 80, 85, 90, 95, 97 or 100% identity to a polynucleotide reference sequence of SEQ ID NO:l, wherein said reference sequence may be identical to the sequence of SEQ ID NO: 1 or may include up to a certain integer number of nucleotide alterations as compared to the reference sequence, wherein said alterations are selected from the group consisting of at least one nucleotide deletion, substitution, including transition and transversion, or insertion, and wherein said alterations may occur at the 5' or 3' terminal positions of the reference nucleotide sequence or anywhere between those terminal positions, interspersed either individually among the nucleotides in the reference sequence or in one or more contiguous groups within the reference sequence, and wherein said number of nucleotide alterations is determined by multiplying the total number of nucleotides in SEQ ID NO: 1 by
  • n n is the number of nucleotide alterations
  • x n is the total number of nucleotides in SEQ ID NO:l
  • y is 0.50 for 50% , 0.60 for 60% , 0.70 for 70%, 0.80 for 80% , 0.85 for 85% , 0.90 for 90%, 0.95 for 95% , 0.97 for 97% or 1.00 for 100%
  • any non- integer product of x n and y is rounded down to the nearest integer prior to subtracting it from x n .
  • Alterations of a polynucleotide sequence encoding the polypeptide of SEQ ID NO:2 may create nonsense, missense or frameshift mutations in this coding sequence and thereby alter the polypeptide encoded by the polynucleotide following such alterations.
  • Preferred polypeptide embodiments further include an isolated polypeptide comprising a polypeptide having at least a 50,60, 70, 80, 85, 90, 95, 97 or 100% identity to a polypeptide reference sequence of SEQ ID NO:2, wherein said reference sequence may be identical to the sequence of SEQ ID NO: 2 or may include up to a certain integer number of am o acid alterations as compared to the reference sequence, wherein said alterations are selected from the group consisting of at least one amino acid deletion, substitution, including conservative and non-conservative substitution, or insertion, and wherein said alterations may occur at the amino- or carboxy-terminal positions of the reference polypeptide sequence or anywhere between those terminal positions, interspersed either individually among the amino acids in the reference sequence or in one or more contiguous groups within the reference sequence, and wherein said number of amino acid alterations is determined by multiplying the total number of amino acids in SEQ ID NO: 2 by the numerical percent of the respective percent identity and subtracting that product from said total number of amino acids in SEQ ID NO:
  • n a is the number of amino acid alterations
  • x a is the total number of amino acids in SEQ ID NO:2
  • y is 0.50 for 50% , 0.60 for 60% , 0.70 for 70% , 0.80 for 80% , 0.85 for 85% , 0.90 for 90% , 0.95 for 95% , 0.97 for 97% or 1.00 for 100% , and wherein any non-integer product of x a and y is rounded down to the nearest integer prior to subtracting it from x,
  • the present invention relates to CBFBJB02 polypeptides (or CBFBJB02 proteins)
  • the CBFBJB02 polypeptides include the polypeptide of SEQ ID NO 2, as well as polypeptides comprising the amino acid sequence of SEQ ID NO 2, and polypeptides comprising the amino acid sequence which have at least 80% identity to that of SEQ ID NO 2 over its entire length, and still more preferably at least 90% identity, and even still more preferably at least 95% identity to SEQ ID NO 2 Furthermore, those with at least 97-99% are highly preferred Also included within CBFBJB02 polypeptides are polypeptides having the ammo acid sequence which have at least 80% identity to the polypeptide having the amino acid sequence of SEQ ID NO 2 over
  • CBFBJB02 polypeptide exhibit at least one biological activity of CBFBJB02
  • the CBFBJB02 polypeptides may be m the form of the "mature" protem or may be a part of a larger protein such as a fusion protein It is often advantageous to include an additional ammo acid sequence which contains secretory or leader sequences, pro-sequences, sequences which aid m purification such as multiple histidme residues, or an additional sequence for stability during recombinant production
  • a fragment is a polypeptide having an amino acid sequence that entirely is the same as part, but not all, of the amino acid sequence of the aforementioned CBFBJB02 polypeptides
  • fragments may be "free-standing,” or comp ⁇ sed within a larger polypeptide of which they form a part or region, most preferably as a single continuous region
  • Representative examples of polypeptide fragments of the invention include, for example, fragments from about amino acid number 1-20, 21-40, 41-60, 61-80, 81-100, and 101 to the end of CBFBJB02 polypeptide
  • “about” includes the particularly recited ranges larger or smaller by several, 5, 4, 3, 2 or 1 amino acid at either extreme or at both extremes
  • Preferred fragments include, for example, truncation polypeptides having the amino acid sequence of CBFBJB02 polypeptides, except for deletion of a continuous senes of residues that includes the amino terminus, or a continuous senes of residues that includes the carboxyl terminus or deletion of two continuous senes of residues, one including the amino terminus and one including the carboxyl terminus
  • fragments characterized by structural or functional att ⁇ butes such as fragments that compnse alpha-helix and alpha-helix forming regions, beta-sheet and beta-sheet-forming regions, turn and turn-forming regions, coil and coil-forming regions, hydrophihc regions, hydrophobic regions, alpha amphipathic regions, beta amphipathic regions, flexible regions, surface-forming regions, substrate binding region, and high antigenic index regions
  • Other preferred fragments are biologically active fragments Biologically active fragments are those that mediate CBFBJB02 activity, including those with a similar activity or an improved activity, or with a
  • the CBFBJB02 polypeptides of the invention can be prepared in any suitable manner.
  • Such polypeptides include isolated natui lly occurring polypeptides, recombinantly produced polypeptides, synthetically produced polypeptides, or polypeptides produced by a combination of these methods. Means for preparing such polypeptides are well understood in the art.
  • CBFBJB02 polynucleotides include isolated polynucleotides which encode the CBFBJB02 polypeptides and fragments, and polynucleotides closely related thereto. More specifically, CBFBJB02 polynucleotide of the invention include a polynucleotide comprising the nucleotide sequence contained in SEQ ID NO: 1 encoding a CBFBJB02 polypeptide of SEQ ID NO: 2, and polynucleotide having the particular sequence of SEQ ID NO: 1.
  • CBFBJB02 polynucleotides further include a polynucleotide comprising a nucleotide sequence that has at least 80% identity over its entire length to a nucleotide sequence encoding the CBFBJB02 polypeptide of SEQ ID NO:2, and a polynucleotide comprising a nucleotide sequence that is at least 80% identical to of SEQ ID NO: 1 over its entire length.
  • polynucleotides at least 90% identical are particularly preferred, and those with at least 95% are especially preferred.
  • those with at least 97% are highly preferred and those with at least 98-99% are most highly preferred, with at least 99% being the most prefeired.
  • CBFBJB02 polynucleotides are also included under CBFBJB02 polynucleotides.
  • a nucleotide sequence which has sufficient identity to a nucleotide sequence contained in SEQ ID NO: 1 to hybridize under conditions useable for amplification or for use as a probe or marker.
  • the invention also provides polynucleotides which are complementary to such CBFBJB02 polynucleotides.
  • CBFBJB02 of the invention is structurally related to other proteins of the DNA binding protein family, as shown by the results of sequencing the cDNA of Table 1 (SEQ ID NOT) encoding human CBFBJB02.
  • the cDNA sequence of SEQ ID NO: 1 contains an open reading frame (nucleotide number 99 to 758) encoding a polypeptide of 220 amino acids of SEQ ID NO:2.
  • the amino acid sequence of Table 2 (SEQ ID NO:2) has about 45.1 % identity (using FASTA) in 184 amino acid residues with mouse chromodoinain-helicase-DNA binding protein CHD-1 (V.Delmas et al.,Proc. Natl. Acad. Sci. U.S.A.
  • CBFBJB02 polypeptides and polynucleotides of the present invention are expected to have, inter alia, similar biological functions/properties to their homologous polypeptides and polynucleotides, and their utility is obvious to anyone skilled in the art.
  • a nucleotide sequence of a human CBFBJB02 (SEQ ID NO 1)
  • One polynucleotide of the present invention encoding CBFBJB02 may be obtained using standard cloning and screening, from a cDNA library denved from mRNA in cells of human cord blood using the expressed sequence tag (EST) analysis (Adams, M D , et al Science (1991) 252 1651- 1656, Adams, M D et al , Nature, (1992) 355 632-634, Adams, M D , et al , Nature (1995) 377 Supp 3-174)
  • Polynucleotides of the invention can also be obtained from natural sources such as genomic DNA libraries or can be synthesized using well known and commercially available techniques
  • the nucleotide sequence encoding CBFBJB02 polypeptide of SEQ ID NO 2 may be identical to the polypeptide encoding sequence contained m Table 1 (nucleotide number 99 to 758 of SEQ ID NO 1), or it may be a sequence, which as a result of the redundancy (degen
  • the polynucleotide may include the coding sequence for the mature
  • the coding sequence for the mature polypeptide or fragment in reading frame with other coding sequences such as those encoding a leader or secretory sequence, a pre-, or pro- or prepro- protein sequence, or other fusion peptide portions
  • a marker sequence which facilitates punfication of the fused polypeptide can be encoded
  • the marker sequence is a hexa-histidine peptide, as provided in the pQE vector (Qiagen, Inc ) and descnbed m Gentz et al , Proc Natl Acad Set USA (1989) 86 821- 824, or is an HA tag
  • the polynucleotide may also contain non-coding 5 ' and 3 ' sequences, such as transcribed, non-translated sequences, splicing and polyadenylation signals, nbosome binding sites and sequences that stabilize mRNA Further preferred embodiments are
  • the present invention further relates to polynucleotides that hybndize to the herein above- desenbed sequences
  • the present mvention especially relates to polynucleotides which hybndize under stringent conditions to the herein above-desenbed polynucleotides
  • stringent conditions means hybndization will occur only if there is at least 80%, and preferably at least 90%, and more preferably at least 95%, yet even more preferably 97-99% identity between the sequences
  • Polynucleotides of the invention which are identical or sufficiently identical to a nucleotide sequence contained m SEQ ID NO 1 or a fragment thereof, may be used as hybndization probes for cDNA and genomic DNA, to isolate full-length cDNAs and genomic clones encoding CBFBJB02 polypeptide and to isolate cDNA and genomic clones of other genes (including genes encoding homologs and orthologs from species other than human) that have a high sequence similanty to the CBFBJB02 gene
  • Such hybndization techniques are known to those of skill in the art
  • these nucleotide sequences are 80% identical, preferably 90% identical, more preferably 95% identical to that of the referent
  • the probes generally will compnse at least 15 nucleotides Preferably, such probes will have at least 30 nucleotides and may have at least 50 nucleotides Particularly preferred probes will range between 30 and 50 nucleotides
  • CBFBJB02 polynucleotides of the present invention further include a nucleotide sequence comprising a nucleotide sequence that hybndize under stringent condition to a
  • CBFBJB02 polypeptides comprising amino acid sequence encoded by nucleotide sequence obtained by the above hybndization condition
  • Stringent hybndization conditions are as defined above or, alternatively, conditions under overnight incubation at 42°C in a solution comp ⁇ sing 50% formamide, 5xSSC (150mM NaCl, 15mM tnsodium citrate), 50 mM sodium phosphate (pH7 6), 5x Denhardt's solution, 10 % dextran sulfate, and 20 microgram/ml denatured, sheared salmon sperm DNA, followed by washing the filters in 0 lx SSC at about 65°C
  • the present invention also relates to vectors which compnse a polynucleotide or polynucleotides of the present invention, and host cells which are genetically engineered with vectors of the invention and to the production of polypeptides of the invention by recombinant techniques Cell-free translation systems can also be employed to produce such proteins using RNAs denved from the DNA constructs of the present invention
  • host cells can be genetically engineered to incorporate expression systems or portions thereof for polynucleotides of the present mvention
  • Introduction of polynucleotides into host cells can be effected by methods desc ⁇ bed in many standard laboratory manuals, such as Davis et al, BASIC METHODS IN MOLECULAR BIOLOGY (1986) and Sambrook et al , MOLECULAR CLONING A LABORATORY MANUAL, 2nd Ed , Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N Y (1989) such as calcium phosphate transfection, DE.AE-dextran mediated transfection, transvection, micro njection, cationic pid-mediated transfection, electroporation, transduction, scrape loading, ballistic introduction or infection
  • appropnate hosts mclude bactenal cells, such as streptococci, staphylococci, E coli, Streptomyces and Bacillus subtihs cells, fiingal cells, such as yeast cells and Aspergillus cells, insect cells such as Drosophila S2 and Spodoptera Sf cells, animal cells such as CHO, COS, HeLa, C127, 3T3, BHK, HEK 293 and Bowes melanoma cells, and plant cells
  • chromosomal, episomal and virus-denved systems e g , vectors denved from bactenal plasmids, from bactenophage, from transposons, from yeast episomes, from insertion elements, from yeast chromosomal elements, from viruses such as baculoviruses, papova viruses, such as SV40, vaccinia viruses, adenoviruses, fo
  • the expression systems may contain control regions that regulate as well as engender expression
  • any system or vector suitable to maintain, propagate or express polynucleotides to produce a polypeptide in a host may be used
  • the appropnate nucleotide sequence may be inserted into an expression system by any of a vanety of well-known and routine techniques, such as, for example, those set forth in Sambrook et al , MOLECULAR CLONING, A LABORATORY MANUAL (supra)
  • approp ⁇ ate secretion signals may be incorporated into the desired polypeptide These signals may be endogenous to the polypeptide or they may be heterologous signals
  • the cells may be harvested p ⁇ or to use m the screening assay
  • the medium can be recovered in order to recover and pu ⁇ fy the polypeptide, if produced intracellularly, the cells must first be lysed before the polypeptide is recovered
  • CBFBJB02 polypeptides can be recovered and pu ⁇ fied from recombinant cell cultures by well-known methods including ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography and lectin chromatography
  • high performance liquid chromatography is employed for pu ⁇ fication
  • Well known techniques for refolding proteins may be employed to regenerate active conformation when
  • This invention also relates to the use of CBFBJB02 polynucleotides for use as diagnostic reagents Detection of a mutated form of CBFB JB02 gene associated with a dysfunction will provide a diagnostic tool that can add to or define a diagnosis of a disease or susceptibility to a disease which results from under-expression, over-expression or altered expression of CBFBJB02 Individuals carrymg mutations in the CBFBJB02 gene may be detected at the DNA level by a vanety of techniques Nucleic acids for diagnosis may be obtained from a subject's cells, such as from blood, urine, saliva, tissue biopsy or autopsy mate ⁇ al The genomic DNA may be used directly for detection or may be amplified enzymatically by using PCR or other amplification techniques p ⁇ or to analysis RNA or cDNA may also be used in similar fashion Deletions and insertions can be detected by a change in size
  • Point mutations can be identified by hyb ⁇ dizing amplified DNA to labeled CBFBJB02 nucleotide sequences Perfectly matched sequences can be distinguished from mismatched duplexes by RNase digestion or by differences in melting temperatures DNA sequence differences may also be detected by alterations in electrophoretic mobility of DNA fragments in gels, with or without denatu ⁇ ng agents, or by direct DNA sequencing See, e g , Myers et al , Science (1985) 230 1242 Sequence changes at specific locations may also be revealed by nuclease protection assays, such as RNase and S 1 protection or the chemical cleavage method See Cotton etal , Proc Natl Acad Sci USA (1985) 85 4397-4401 In another embodiment, an array of ohgonucleotides probes comp ⁇ sing CBFBJB02 nucleotide sequence or fragments thereof can be constructed to conduct efficient screening of e g , genetic mutation
  • the diagnostic assays offer a process for diagnosing or determining a susceptibility to cancer, autoimmune disease, and AIDS through detection of mutation in the CBFBJB02 gene by the methods desc ⁇ bed
  • cancer, autoimmune disease, and AIDS can be diagnosed by methods comprising determining from a sample denved from a subject an abnormally decreased or increased level of CBFBJB02 polypeptide or CBFBJB02 mRNA Decreased or mcreased expression can be measured at the RNA level usmg any of the methods well known in the art for the quantitation of polynucleotides, such as, for example, PCR, RT-PCR, RNase protection, Northern blotting and other hybndization methods ⁇ say techniques that can be used to determine levels of a protein, such as an CBFBJB02 polypeptide, in a sample denved from a host are well-known to those of skill in the art Such assay methods include radioimmunoassays, competitive-binding assays, Western Blot analysis and ELISA assays
  • the present invention relates to a diagonostic kit for a disease or suspectabihty to a disease, particularly cancer, autoimmune disease, and AIDS, which comprises (a) a CBFBJB02 polynucleotide, preferably the nucleotide sequence of SEQ ID NO 1, or a fragment thereof, (b) a nucleotide sequence complementary to that of (a),
  • a CBFBJB02 polypeptide preferably the polypeptide of SEQ ID NO 2, or a fragment thereof, or
  • any such kit, (a), (b), (c) or (d) may comp ⁇ se a substantial component
  • the nucleotide sequences of the present invention are also valuable for chromosome identification
  • the sequence is specifically targeted to and can hybndize with a particular location on an individual human chromosome
  • the mapping of relevant sequences to chromosomes according to the present invention is an important first step m correlating those sequences with gene associated disease
  • genetic map data are found, for example, in V McKusick, Mendelian Inhe ⁇ tance in Man (available on line through Johns Hopkins University Welch Medical Library)
  • the relationship between genes and diseases that have been mapped to the same chromosomal region are then identified through linkage analysis (coinhe ⁇ tance of physically adjacent genes)
  • the differences m the cDNA or genomic sequence between affected and unaffected individuals can also be determined If a mutation is observed in some or all of the affected individuals but not in any normal individuals
  • polypeptides of the invention or their fragments or analogs thereof, or cells expressing them can also be used as l munogens to produce antibodies lmmunospecific for the CBFBJB02 polypeptides
  • the term "lmmunospecific" means that the antibodies have substantial!
  • Antibodies generated against the CBFBJB02 polypeptides can be obtained by administering the polypeptides or epitope-bea ⁇ ng fragments, analogs or cells to an animal, preferably a nonhuman, using routine protocols For preparation of monoclonal antibodies, any technique which provides antibodies produced by continuous cell lme cultures can be used Examples include the hyb ⁇ doma technique (Kohler, G and Mdstein, C , Nature (1975) 256 495-497), the t ⁇ oma technique, the human B-cell hyb ⁇ doma technique (Kozbor et al , Immunology Today (1983) 4 72) and the EBV-hyb ⁇ doma technique (Cole et al , MONOCLONAL ANTIBODIES AND CANCER THERAPY, pp 77-96, Alan R L ⁇ ss, Inc , 1985)
  • the above-desc ⁇ bed antibodies may be employed to isolate or to identify clones expressing the polypeptide or to pu ⁇ fy the polypeptides by affinity chromatography
  • Antibodies against CBFBJB02 polypeptides may also be employed to treat cancer, autoimmune disease, and ./MDS, among others
  • Vaccines Another aspect of the invention relates to a method for inducing an immunological response in a mammal which comprises inoculating the mammal with CBFBJB02 polypeptide, or a fragment thereof, adequate to produce antibody and/or T cell immune response to protect said animal from cancer, autoimmune disease, and AIDS, among others
  • a method of inducing immunological response m a mammal which comprises, dehvenng CBFBJB02 polypeptide via a vector directmg expression of CBFBJB02 polynucleotide in vivo in order to induce such an immunological response to produce antibody to protect said animal from diseases
  • composition which, when introduced into a mammalian host, induces an immunological response in that mammal to a CBFBJB02 polypeptide wherein the composition composes a CBFB.IB02 polypeptide or CBFBJB02 gene
  • the vaccine formulation may further comprise a suitable carrier Since CBFBJB02 polypeptide may be broken down in the stomach, it is preferably administered parenterally (including subcutaneous, intramuscular, intravenous, intradermal etc injection)
  • parenterally including subcutaneous, intramuscular, intravenous, intradermal etc injection
  • Formulations suitable for parenteral administration include aqueous and non-aqueous stenle mjection solutions which may contain anti-oxidants, buffers, bacte ⁇ ostats and solutes which render the formulation mstonic with the blood of the recipient, and aqueous and non-aqueous sterile suspensions which may include suspending agents or thickening agents
  • the formulations may be presented in unit-dose or multi-dose containers, for
  • the CBFBJB02 polypeptide of the present invention may be employed in a screening process for compounds which activate (agonists) or inhibit activation of (antagonists, or otherwise called inhibitors) the CBFBJB02 polypeptide of the present invention
  • polypeptides of the mvention may also be used to assess identify agonist or antagonists from, for example, cells, cell-free preparations, chemical libranes, and natural product mixtures
  • agonists or antagonists may be natural or modified substrates, ligands, receptors, enzymes, etc , as the case may be, of the polypeptide of the present invention, or may be structuial or functional mimetics of the polypeptide of the present mvention See Cohgan etal , Current Protocols in Immunology 1(2) Chapter 5 (1991) CBFBJB02 polypeptides are responsible for many biological functions, mcluding many pathologies
  • agonists are employed for therapeutic and prophylactic purposes for such conditions as cancer, autoimmune disease, and AIDS Antagonists may be employed for a vanety of therapeutic and prophylactic purposes for such conditions as cancer, autoimmune disease, and AIDS
  • such screening procedures may involve using appropnate cells which express the CBFBJB02 polypeptide or respond to CBFBJB02 polypeptide of the present invention
  • Such cells include cells from mammals, yeast, Drosophila or E coli Cells which express the CBFBJB02 polypeptide (or cell membrane containing the expressed polypeptide) or respond to CBFBJB02 polypeptide are then contacted with a test compound to observe bmdmg, or stimulation or inhibition of a functional response The ability of the cells which were contacted with the candidate compounds is compared with the same cells which were not contacted for CBFBJB02 activity
  • the assays may simply test bmdmg of a candidate compound wherein adherence to the cells bearing the CBFBJB02 polypeptide is detected by means of a label directly or indirectly associated with the candidate compound or in an assay involving competition with a labeled competitor Further, these assays may test whether the candidate compound results in a signal generated by activation of the CBFBJB02 polypeptide, using detection systems approp ⁇ ate to the cells beanng the CBFBJB02 polypeptide Inhibitors of activation are generally assayed in the presence of a known agonist and the effect on activation by the agomst by the presence of the candidate compound is observed
  • the assays may simply comprise the steps of mixing a candidate compound with a solution containing a CBFBJB02 polypeptide to form a mixture, measuring CBFBJB02 activity in the mixture, and comparing the CBFBJB02 activity of the mixture to a standard
  • the CBFBJB02 cDNA, protein and antibodies to the protein may also be used to configure assays for detecting the effect of added compounds on the production of CBFBJB02 mRNA and protem m cells
  • an ELISA may be constructed for measuring secreted or cell associated levels of CBFBJB02 protem using monoclonal and polyclonal antibodies by standard methods known in the art, and this can be used to discover agents which may inhibit or enhance the production of CBFBJB02 (also called antagonist or agonist, respectively) from suitably manipulated cells or tissues
  • the CBFBJB02 protem may be used to identify membrane bound or soluble receptors, if any, through standard receptor binding techniques known m the art These include, but are not limited to, hgand binding and crosshnkmg assays in which the CBFBJB02 is labeled with a radioactive isotope (eg 1251), chemically modified (eg biotinylated), or fused to a peptide sequence suitable for detection or punfication, and incubated with a source of the putative receptor (cells, cell membranes, cell supernatants, tissue extracts, bodily fluids) Other methods include biophysical techniques such as surface plasmon resonance and spectroscopy In addition to bemg used for punfication and cloning of the receptor, these binding assays can be used to identify agonists and antagonists of CBFBJB02 which compete with the binding of CBFBJB02 to its receptors, if any Standard methods for conducting screening assays are well understood in the art Examples of potential CB
  • the present invention relates to a screening kit for identifying agonists, antagonists, ligands, receptors, substrates, enzymes, etc for CBFBJB02 polypeptides, or compounds which decrease or enhance the production of CBFBJB02 polypeptides, which comprises
  • This invention provides methods of treating abnormal conditions such as, cancer, autoimmune disease, and AIDS, related to both an excess of and insufficient amounts of CBFBJB02 polypeptide activity
  • CBFBJB02 polypeptide If the activity of CBFBJB02 polypeptide is in excess, several approaches are available One approach compnses administering to a subject an inhibitor compound (antagonist) as hereinabove desc ⁇ bed along with a pharmaceutically acceptable earner m an amount effective to inhibit the function of the CBFBJB02 polypeptide, such as, for example, by blocking the binding of ligands, substrates, receptors, enzymes, etc , or by inhibiting a second signal, and thereby alleviating the abnormal condition. In another approach, soluble forms of CBFBJB02 polypeptides still capable of binding the hgand, substrate, enzymes, receptors, etc m competition with endogenous CBFBJB02 polypeptide may be administered Typical embodiments of such competitors compnse fragments of the CBFBJB02 polypeptide
  • expression of the gene encoding endogenous CBFBJB02 polypeptide can be inhibited using expression blocking techniques
  • Known such techniques involve the use of antisense sequences, either internally generated or separately administered See, for example, O'Connor, J Neurochem (1991) 56 560 in Ohgodeoxynucleotides as Antisense Inhibitors of Gene Expression, CRC Press, Boca Raton, FL (1988)
  • ohgonucleotides which form t ⁇ ple helices with the gene can be supplied See, for example, Lee et al , Nucleic Acids Res (1979) 6 3073, Cooney et al , Science (1988) 241 456, Dervan et ⁇ / , Science (1991) 251 1360
  • These ohgomers can be administered per se or the relevant o gomers can be expressed in vivo
  • a polynucleotide of the mvention may be engmeered for expression m a replication defective retroviral vector, as discussed above
  • the retroviral expression construct may then be isolated and introduced into a packagmg cell transduced with a retroviral plasrmd vector containing RNA encoding a polypeptide of the present mvention such that the packagmg cell now produces infectious viral particles containing the gene of interest
  • These producer cells may be administered to a subject for engineering cells in vivo and
  • Peptides such as the soluble form of CBFBJB02 polypeptides, and agonists and antagonist peptides or small molecules, may be formulated in combination with a suitable pharmaceutical earner
  • a suitable pharmaceutical earner Such formulations compnse a therapeutically effective amount of the polypeptide or compound, and a pharmaceutically acceptable earner or excipient
  • earners include but are not limited to, saline, buffered saline, dextrose, water, glycerol, ethanol, and combinations thereof Formulation should suit the mode of administration, and is well within the skill of the art
  • the mvention further relates to pharmaceutical packs and kits compnsing one or more containers filled with one or more of the ingredients of the aforementioned compositions of the mvention
  • Polypeptides and other compounds of the present mvention may be employed alone or in conjunction with other compounds, such as therapeutic compounds
  • Preferred forms of systemic administration of the pharmaceutical compositions include injection, typically by intravenous mjection Other mjection routes, such as subcutaneous, intramuscular, or intrape ⁇ toneal, can be used
  • Alternative means for systemic administration include transmucosal and transdermal administration using penetrants such as bile salts or fusidic acids or other detergents
  • oral administration may also be possible
  • Administration of these compounds may also be topical and/or localized, in the form of salves, pastes, gels and the like
  • Suitable dosages are in the range of 0 1-100 ⁇ g/kg of subject Wide vanations in the needed dosage, however, are to be expected in view of the vanety of compounds available and the differing efficiencies of va ⁇ ous routes of administration
  • oral administration would be expected to require higher dosages than administration by intravenous mjection Vanations m
  • these dosage levels can be adjusted usmg standard empi ⁇ cal routines for optim-zation, as is well understood in the art
  • Polypeptides used in treatment can also be generated endogenously in the subject, in treatment modalities often refened to as "gene therapy" as desc ⁇ bed above
  • cells from a subject may be engineered with a polynucleotide, such as a DNA or RNA, to encode a polypeptide ex vivo, and for example, by the use of a retroviral plasm

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Toxicology (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention concerne des polypeptides et des polynucléotides CBFBJB02 et des procédés d'obtention de ces polypeptides par des techniques recombinantes. Font aussi l'objet de cette invention des procédés d'utilisation des polypeptides et des polynucléotides CBFBJB02 dans la conception de protocoles pour le traitement de cancers, de maladies auto-immunes et du SIDA, entre autres, et des doses diagnostiques pour ces pathologies.
PCT/CN1998/000036 1998-03-12 1998-03-12 Gene humain de type chd-1 Ceased WO1999046294A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/CN1998/000036 WO1999046294A1 (fr) 1998-03-12 1998-03-12 Gene humain de type chd-1

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/CN1998/000036 WO1999046294A1 (fr) 1998-03-12 1998-03-12 Gene humain de type chd-1

Publications (1)

Publication Number Publication Date
WO1999046294A1 true WO1999046294A1 (fr) 1999-09-16

Family

ID=4575031

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN1998/000036 Ceased WO1999046294A1 (fr) 1998-03-12 1998-03-12 Gene humain de type chd-1

Country Status (1)

Country Link
WO (1) WO1999046294A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1402053A4 (fr) * 2001-06-05 2005-05-11 Exelixis Inc Chds en tant que modulateurs du mecanisme d'action de p53 et utilisations

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
DEV. BIOL., 161(1), (1994), RANDAZZO F.M. et al., "Brg1: A Putative Murine Homologue of the Drosophila Brahma Gene, a Homeotic Gene Regulator", pages 229-242. *
EMB, X97796, (1996), KANAAR R. et al., "M. Musculus mRNA Homologous to S. Cerevisiae RAD54". *
NUCLEIC ACIDS RES., 22(10), (1994), CHIBA H. et al., "Two Human Homologues of Saccharomyces Cerevisiae SW12/SNF2 and Drosophila Brahma are Transcriptional Coactivators Cooperating with the Estrogen Receptor and Retinoic Acid Receptor", pages 1815-1820. *
PROC. NATL. ACAD. SCI. U.S.A., 90, (1993), ANDEYSON S.K. et al., "A Cyclophilin-Related protein Involved in the Function of Natural Killer Cells", pages 542-546. *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1402053A4 (fr) * 2001-06-05 2005-05-11 Exelixis Inc Chds en tant que modulateurs du mecanisme d'action de p53 et utilisations

Similar Documents

Publication Publication Date Title
US6319688B1 (en) Polynucleotide encoding human sodium dependent phosphate transporter (IPT-1)
WO1999021991A1 (fr) Bmzf12: gene a doigt de zinc clone a partir de la moelle osseuse
WO1999046293A1 (fr) Proteine a doigt de zinc derivee de cellules hematopoietiques
US5952483A (en) Human IκB-β
EP0908515A2 (fr) Polypeptide pancréatique
EP0875572A2 (fr) CRFG-1a, cible et marqueur pour l'insuffisance rénale chronique
WO1999046295A1 (fr) Gene homologue au gene b(2)gene de drosophile et proteine ypr015c de levure putative 26,5kd
WO1999036526A1 (fr) Cbmajc02: gene semblable au complexe de synthase bovine f1f0-atp de f-sous-unite a membrane f¿0?
WO1999046294A1 (fr) Gene humain de type chd-1
WO1999046290A1 (fr) Gene humain de type recepteur d'angiotensine ii/vasopreessine (aii/avp) (cbdakd01)
EP1224220A2 (fr) Gene 1 de reponse primaire a l'erythropoietine (epo), gene eprg1
WO1999022006A1 (fr) Cblafc02: sous-unite de h(+)-atpase vacuolaire
WO1999021885A1 (fr) Gene transporteur 7 abc humain (habc7)
WO1999022007A1 (fr) Cbfaie10: gene skd1 humain
WO1999036527A1 (fr) Gene de type ataxine 2
WO1999046291A1 (fr) Gene homologue humain du facteur de maturation des cellules gliales (gmf) type beta (cbfboe11)
WO1999047664A1 (fr) Gene humain homologue du proteolipide mitochondrial mp68: cbnaac01
WO1999047667A1 (fr) Gene humain de sous-unite gamma sec61: cbdame07
EP0892050A2 (fr) HFIZG53 humaine
WO1999046292A1 (fr) Gene humain p18 (cbdara04)
WO1999047559A1 (fr) Isoforme 1 de gene de type dimethylglycine dehydrogenase
WO1999021982A1 (fr) Gene m6b1 humain
EP0889126A2 (fr) Polypeptide YNL075W/HTXFT19
WO1999036525A1 (fr) Cbfblh12: gene fortement associe au gene ci-kfyi bovin pour complexe d'ubiquinone oxyreductase
WO1999021986A1 (fr) Cbfbga09: homologue sl15 humain

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): CN JP US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE

121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: 09091815

Country of ref document: US

NENP Non-entry into the national phase

Ref country code: CA

122 Ep: pct application non-entry in european phase