WO1999036527A1 - An ataxin-2 like gene - Google Patents

Abstract

Ataxin-2 like gene polypeptides and polynucleotides and methods for producing such polypeptides by recombinant techniques are disclosed. Also disclosed are methods for utilizing ataxin-2 like gene polypeptides and polynucleotides in the design of protocols for the treatment of ataxia, cardiomyopathy, deafness, neurological disease, cancer, and AIDS, among others, and diagnostic assays for such conditions.

Description

An Ataxin-2 Like Gene

FIELD OF INVENTION

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 ataxin-2 family, hereinafter referred to as ataxin-2 like gene. The invention also relates to inhibiting or activating the action of such polynucleotides and polypeptides.

BACKGROUND OF THE INVENTION

Ataxin-2 is the gene responsible for spinocerebellar ataxia, type 2. A partial cDNA sequence obtained by Pulst was found to be homologous to ataxin-2.

This indicates that the ataxin-2 family has an established, proven history as therapeutic targets. Clearly there is a need for identification and characterization of further members of the ataxin-2 family which can play a role in preventing, ameliorating or correcting dysfunctions or diseases, including, but not limited to, ataxia, cardiomyopathy, deafness, neurological disease, cancer, and AIDS.

SUMMARY OF THE INVENTION In one aspect, the invention relates to ataxin-2 like gene polypeptides and recombinant materials and methods for their production. Another aspect of the invention relates to methods for using such ataxin-2 like gene polypeptides and polynucleotides. Such uses include the treatment of ataxia, cardiomyopathy, deafness, neurological disease, cancer, and AIDS, among others. In still another aspect, the invention relates to methods to identify agonists and antagonists using the materials provided by the invention, and treating conditions associated with ataxin-2 like gene imbalance with the identified compounds. Yet another aspect of the invention relates to diagnostic assays for detecting diseases associated with inappropriate ataxin-2 like gene activity or levels.

DESCRIPTION OF THE INVENTION Definitions

The following definitions are provided to facilitate understanding of certain terms used frequently herein.

"Ataxin-2 like gene" refers, among others, generally to a polypeptide having the ammo acid sequence set forth in SEQ ID NO:2 or an allelic variant thereof. "Ataxιn-2 like gene activity or ataxιn-2 like gene polypepϋde activity" or "biological activity of the ataxιn-2 like gene or ataxιn-2 like gene polypeptide" refers to the metabolic or physiologic function of said ataxιn-2 like gene including similar acuvities or improved activities or these activities with decreased undesirable side-effects Also mcluded are antigemc and immunogenic activities of said ataxιn-2 like gene

"Ataxιn-2 like 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 mcludes polyclonal and monoclonal antibodies, chimenc, single chain, and humanized antibodies, as well as Fab fragments, including the products of an Fab or other immunoglobulin 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 in a living animal is not "isolated," but the same polynucleotide or polypeptide separated from the coexisting materials of its natural state is "isolated", as the term is employed herein

"Polynucleotide" generally refers to any polynbonucleotide or polydeoxnbonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA "Polynucleotides" mclude, without limitation single- and double-stranded DNA, DNA that is a mixture of single- and double- stranded regions, smgle- and double-stranded RNA, and RNA that is mixture of single- and double-stranded regions, hybπd molecules compπsmg DNA and RNA that may be smgle-stranded or, more typically, double-stranded or a mixture of smgle- and double-stranded regions In addition, "'polynucleotide" refers to tπple-stranded regions compπsmg RNA or DNA or both RNA and DNA The term polynucleotide also mcludes DNAs or RNAs containing one or more modified bases and DNAs or RNAs with backbones modified for stability or for other reasons "Modified" bases mclude, for example, tπtylated bases and unusual bases such as inosine A vaπety of modifications has been made to DNA and RNA, thus, "polynucleotide" embraces chemically, enzymatically or metabo cally modified forms of polynucleotides as typically found in nature, as well as the chemical forms of DNA and RNA characteπstic of viruses and cells "Polynucleotide" also embraces relatively short polynucleotides. often referred to as oligonucleotides "Polypeptide" refers to any peptide or protein compπsmg two or more ammo acids jomed to each other by peptide bonds or modified peptide bonds, l e , peptide isosteres "Polypeptide" refers to both short chains, commonly referred to as peptides, ohgopeptides or o gomers, and to longer chains, generally referred to as proteins Polypeptides may contain ammo acids other than the 20 gene-encoded ammo acids "Polypeptides" mclude ammo acid sequences modified either by natural processes, such as posttranslational processmg, or bv chemical modification techniques which are well known in the art Such modifications are well descπbed m basic texts and in more detailed monographs, as well as in a voluminous research literature Modifications can occur anywhere m a polypeptide, mcludmg the peptide backbone, the ammo acid side-chains and the ammo or carboxyl termini It will be appreciated that the same type of modification may be present in the same or varying degrees at several sites in a given polypeptide Also, a given polypeptide may contain many types of modifications Polypeptides may be branched as a result of ubiquiunation, and they may be cyclic, with or without branching Cyclic, branched and branched cyclic polypeptides may result from posttranslation natural processes or may be made by synthetic methods Modifications mclude acetylation, acylation, ADP-πbosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide denvanve, covalent attachment of a lipid or pid deπvative, covalent attachment of phosphotidylinositol, cross-linking, cychzation, disulfide bond formation, demethvlation, formation of covalent cross-links, formation of cystine, formation of pyroglutamate, formylation, gamma- carboxylation, glycosylation, GPI anchor formation, hydroxylation. lodination, methylation, myπstoylation, oxidation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation. sulfation, transfer-RNA mediated addition of a mo acids to proteins such as arginylation, and ubiquiunation See, for instance, PROTEINS - STRUCTURE AND MOLECULAR PROPERTIES, 2nd Ed , T E Creιghton, W H Freeman and Company, New York, 1993 and Wold, F , Posttranslational Protem Modifications Perspectives and Prospects, pgs 1-12 m POSTTRANSLATIONAL COVALENT MODIFICATION OF PROTEINS, B C Johnson, Ed , Academic Press, New York. 1983. Seifter et al , "Analysis for protem modifications and nonprotein cofactors", Meth Enzymol (1990) 182 626-646 and Rattan et al , "Protem Synthesis Posttranslational Modifications and Aging", Ann NYAcadSci (1992) 663 48-62 "Vaπant" as the term is used herein, is a polynucleotide or polypeptide that differs from a reference polynucleotide or polypeptide respectively, but retains essential properties A typical vaπant of a polynucleotide differs m nucleotide sequence from another, reference polynucleotide Changes in the nucleotide sequence of the vaπant may or may not alter the ammo acid sequence of a polypeptide encoded by the reference polynucleotide Nucleotide changes may result in ammo acid substitutions, additions, deletions, fusions and truncations in the polypeptide encoded by the reference sequence, as discussed below A typical vaπant of a polypeptide differs m ammo acid sequence from another, reference polypeptide Generally, differences are limited so that the sequences of the reference polypeptide and the vaπant are closely similar overall and, in many regions, identical A vaπant and reference polypeptide may differ in ammo acid sequence by one or more substitutions, additions, deletions in any combination. 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 measure of the identity of nucleotide sequences or amino acid sequences. In general, the sequences are aligned so that the highest order match is obtained. "Identity" per se has an art-recognized meaning and can be calculated using published techniques. See, e.g.: (COMPUTATIONAL MOLECULAR BIOLOGY, Lesk, A.M., ed., Oxford University Press, New York, 1988; BIOCOMPUTING: INFORMAΗCS AND GENOME PROJECTS, Smith, D.W., ed., Academic Press, New York, 1993; COMPUTER ANALYSIS OF SEQUENCE DATA, PART I, Griffin, A.M., and Griffin, H.G., eds., Humana Press, New Jersey, 1994; SEQUENCE ANALYSIS IN MOLECULAR BIOLOGY, von Heinje, G., Academic Press, 1987; and SEQUENCE ANALYSIS PRIMER, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991). While there exist a number of methods to measure identity between two polynucleotide or polypeptide sequences, the term "identity" is well known to skilled artisans (Carillo, H., and Lipton, D., SIAM J Applied Math (1988) 48:1073). Methods commonly employed to determine identity or similarity between two sequences include, but are not limited to, those disclosed in Guide to Huge Computers, Martin J. Bishop, ed., Academic Press, San Diego, 1994, and Carillo, H., and Lipton, D., SIAM J Applied Math (1988) 48:1073. Methods to determine identity and similarity are codified in computer programs. Preferred computer program methods to determine identity and similarity between two sequences include, but are not limited to, GCS program package (Devereux, J., et al., Nucleic Acids Research (1984) 12(1):387), BLASTP, BLASTN, FASTA (Atschul, S.F. et al., JMolec Biol (1990) 215:403). As an illustration, by a polynucleotide having a nucleotide sequence having at least, for example, 95% "identity" to a reference nucleotide sequence of SEQ ID NO: 1 is intended that the nucleotide sequence of the polynucleotide is identical to the reference sequence except that the polynucleotide sequence may include up to five point mutations per each 100 nucleotides of the reference nucleotide sequence of SEQ ID NO: 1. In other words, to obtain a polynucleotide having a nucleotide sequence at least 95% identical to a reference nucleotide sequence, up to 5% of the nucleotides in the reference sequence may be deleted or substituted with another nucleotide, or a number of nucleotides up to 5% of the total nucleotides in the reference sequence may be inserted into the reference sequence. These mutations of the reference sequence may occur at the 5 or 3 terminal positions of the reference nucleotide sequence or anywhere between those terminal positions, interspersed either individually among nucleotides in the reference sequence or in one or more contiguous groups within the reference sequence.

Similarly, by a polypeptide having an amino acid sequence having at least, for example, 95% "identity" to a reference amino acid sequence of SEQ ED NO:2 is intended that the amino acid sequence of the polypeptide is identical to the reference sequence except that the polypeptide sequence may include up to five amino acid alterations per each 100 amino acids of the reference amino acid of SEQ ID NO: 2. In other words, to obtain a polypeptide having an amino acid sequence at least 95% identical to a reference amino acid sequence, up to 5% of the amino acid residues in the reference sequence may be deleted or substituted with another amino acid, or a number of amino acids up to 5% of the total amino acid residues in the reference sequence may be inserted into the reference sequence. These alterations of the reference sequence may occur at the amino or carboxy terminal positions of the reference amino acid sequence or anywhere between those terminal positions, interspersed either individually among residues in the reference sequence or in one or more contiguous groups within the reference sequence.

Polypeptides of the Invention

In one aspect, the present invention relates to ataxin-2 like gene polypeptides (or ataxin-2 like gene proteins). The ataxin-2 like gene 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 ataxin-2 like gene polypeptides are polypeptides having the amino acid sequence which have at least 80% identity to the polypeptide having the amino acid sequence of SEQ ID NO:2 over its entire length, and still more preferably at least 90% identity, and still more preferably at least 95% identity to SEQ ID NO:2. Furthermore, those with at least 97-99% are highly preferred. Preferably, ataxin-2 like gene polypeptides exhibit at least one biological activity of ataxin-2 like gene.

The ataxin-2 like gene polypeptides may be in the form of the "mature" protein or may be a part of a larger protein such as a fusion protein. It is often advantageous to include an additional amino acid sequence which contains secretory or leader sequences, pro-sequences, sequences which aid in purification such as multiple histidine residues, or an additional sequence for stability during recombinant production. Fragments of the ataxin-2 like gene polypeptides are also included in the invention. 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 ataxin-2 like gene polypeptides. As with ataxin-2 like gene polypeptides, fragments may be "free-standing," or comprised 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 the ataxin-2 like gene polypeptide. In this context "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 ataxin-2 like gene polypeptides, except for deletion of a continuous series of residues that includes the amino terminus, or a continuous series of residues that includes the carboxyl terminus or deletion of two continuous series of residues, one including the amino terminus and one including the carboxyl terminus. Also preferred are fragments characterized by structural or functional attributes such as fragments that comprise alpha-helix and alpha-helix forming regions, beta-sheet and beta-sheet- foiming regions, turn andtum-forming regions, coil and coil-forming regions, hydrophilic 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 ataxin-2 like gene activity, including those with a similar activity or an improved activity, or with a decreased undesirable activity. Also included are those that are antigenic or immunogenic in an animal, especially in a human.

Preferably, all of these polypeptide fragments retain the biological activity of the ataxin-2 like gene, including antigenic activity. Variants of the defined sequence and fragments also form part of the present invention. Preferred variants are those that vary from the referents by conservative amino acid substitutions - i.e., those that substitute a residue with another of like characteristics. Typical such substitutions are among Ala, Val, Leu and He; among Ser and Thr; among the acidic residues Asp and Glu; among Asn and Gin; and among the basic residues Lys and Arg; or aromatic residues Phe and Tyr. Particularly preferred are variants in which several, 5-10, 1-5, or 1-2 amino acids are substituted, deleted, or added in any combination. The ataxin-2 like gene polypeptides of the invention can be prepared in any suitable manner.

Such polypeptides include isolated naturally 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. Polynucleotides of the Invention

Another aspect of the mvention relates to ataxιn-2 like gene polynucleotides. Ataxιn-2 like gene polynucleotides mclude isolated polynucleotides which encode the ataxιn-2 like gene polypeptides and fragments, and polynucleotides closely related thereto More specifically, ataxιn-2 like gene polynucleotides of the mvention mclude a polynucleotide compπsmg the nucleotide sequence contained in SEQ ID NO 1 encoding an ataxιn-2 like gene polypeptide of SEQ ED NO 2, and polynucleotide having the particular sequence of SEQ ID NO 1 Ataxιn-2 like gene polynucleotides further mclude a polynucleotide compπsmg a nucleotide sequence that has at least 80% identity over its entire length to a nucleotide sequence encoding the alaxιn-2 like gene polypeptide of SEQ ID NO .2, and a polynucleotide compπsmg a nucleotide sequence that is at least 80% identical to of SEQ ID NO 1 over its entire length In this regard, polynucleotides at least 90% identical are particularly preferred, and those with at least 95% are especially preferred Furthermore, 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 preferred Also mcluded under ataxιn-2 like gene polynucleotides are a nucleotide sequence which has sufficient identity to a nucleotide sequence contamed in SEQ ID NO 1 to hybπdize under conditions useable for amplification or for use as a probe or marker The mvention also provides polynucleotides which are complementary to such ataxιn-2 like gene polynucleotides

The atax -2 like gene of the mvention is structurally related to other proteins of the ataxιn-2 family, as shown by the results of sequencing the cDNA of Table 1 (SEQ ID NO 1) encoding human ataxιn-2 like gene The cDNA sequence of SEQ ID NO 1 contains an open reading frame (nucleotide number 63 to 3215) encoding a polypeptide of 1051 ammo acids of SEQ ID NO 2. The amino acid sequence of Table 2 (SEQ ID NO 2) has about 54 3% identity (using Bestfit of GCG) in 342 ammo acid residues with ataxιn-2 (S M Pulst et al Nat Genet 14269-276, 1996) The nucleotide sequence of Table 1 (SEQ ID NO 1) has about 62 7% identity (using Bestfit of GCG) in 1305 nucleotide residues with ataxιn-2 (S M Pulst et al Nat Genet 14269-276, 1996) Thus, ataxιn-2 like gene polypeptides and polynucleotides of the present mvention are expected to have, inter a a, similar biological functions/properties to their homologous polypeptides and polynucleotides, and their utility is obvious to anyone skilled in the art

Table 1' cgggacgcrcctcacttcctagatgggatggcggccgggaagaggcgctcctcatttcctagatgggatggcggccgggc agagatgctcctcactttcctccagccgcgagaccccctccccttccgcctcgcggcgcttcctcgcgccgcggttttcr ctctccacccccgacaccgcggggctccccccgcccgcccacggcgggccccggctgcccgatccccctcgcttcccgcg ctctccagcggggccccagccccggccccctcrctccctcccttctctctaattccccttccggacgctgccatcatgtt gaagccrcagccgctacaacagccctcccaσccccagcagccqccccccacgcaacatgccgtggcccgtcggccccccg gggσcacaαcccrcccaacσgcqgccτcccσαgαccαctgσccacctcταcggcrcc cccqσαccτccaqcσgccαcct ccccctgcctggggcctgtggccgctgccgggagcgggctccgccggggagccgaaggcatcttggcgccgcagccgccg ccgccgcagcaacaccaggagaggccgggggcagccgccatcggcagcgccaggggacagagcacaggaaagggaccccc acagtcacctgtgtttgaaggcgtctacaacaat-ccagaatgctgcatttcct acagctgttgtgggctccactrgtg a gtaaaggtgaaaaatggtaccactta gagggtatcttcaagacgccaagctcaaagtttgaactagccgtggatgct gtgcaccggaaagcatctgagccagcaggtggccctcgtcgggaggacattgtggacaccatggtgtttaagccaagtga tgtcatgcttgttcacttccgaaatgttgacttcaactargc actaaagacaagttcaccgatrcagccattgccatga ac cgaaagtgaatggggaacacaaagagaaggtgcrtcagcgctgggaggggggtgacagcaacagcgacgactatgac ctcgagtctgacatgtccaatggatgggaccccaatgaaatgttcaagttcaatgaggagaactacggtgtgaagactac ctatgatagcagtctttcttcttatacggtgcccttagaaaaggacaactcagaagagtttcgtcagcgagagctgcgtg cggcccagttggctcgagagattgaatcaagcccccagtaccgcctacggatcgccatggagaacgacgatgggcgcact gaaςaggagaagcacagtgcagtccagcggcagggctcagggcgggagagccccagcttggcatccagggaggggaagta tatccctctgcctcaacgagtccgggaaggtccccggggaggagttcgatgcagcagcrctcggggcggtcggcctggcc ttagctctttgccacctcgtggccctcaccatctggacaacagcagccctggcccaggttctgaggcccgtggtatcaat ggaggcccttcccgcatgtccccaaaggcacagcggcctctgagaggtgccaagactc gtcttcgcccagtaataggct tctggagaacttctgttccacctcctcctgcagtgggccggatgtatccccgcgttcrcccaagtctgctgcccctgccc caatctcagcttcctgtccagagcctcccatcggctcggcagtgccaacctcttcagcctccatccctgtgacctcatca gtctcagatcctggagtgggctccatttctccagcttctccaaagatctccctggcccccacagatgtaaaagaac ctc taccaaggaacctgggagaactctggagccccaggagctggctcggatagctgggaaagtccctggtcttcagaatgaac agaaacgattccaactggaagaactgagaaagtttggggcccagtttaagctrcagcccagtagctcccctgagaacagc ctggatccttttcctccccggatcttaaaggaggagcccaaaggaaaggagaaagaggttgatggtctgttgacttcaga gcccatggggtctcccgtctcctccaagacagagtccgtatcggataaggaggacaaaccacccctggcaccatcaggag gcactgaggggccagagcagcccccaccaccttgtccaagccaaactggcagccccccggtgggcctcatcaagggagaa gacaaagatgagggccctgttgctgaacaagtaaagaaatcaacgttgaacccraatgctaaggagttcaatcctacaaa gcctctgctgtctgtgaataaatccaccagtaccccaacttctccggggccccggactcattcaactccctccatcccgg tgctgacagcaggccagagtgggctatacagcccccagtacatctccracatacctcagatccacatgggaccagctgtg caggcacctcagatgta ccatatcctgtatccaattcagtgcctgggcagcagggcaagtaccggggagcaaaaggctc ccttcctccgcagcgcrcggaccaacaccagccagcctcagccccgccgatgatgcaggccgccgcggctgctggcccgc ctctggtggccgccacgccccattcrtcctacatccccracaacccrcagcagttcccaggccagccagccatgatgcag cccatggcccacraccccTicacagccggtgrtrgcccccatgcttcagagcaacccacgcatgctgacgtcgggcagcca -ccccaggccatcgtgtcatcctctacccctcaaτacccttcrgcagagcagcctaccccccaagccctt atgccactg ttcaccagtcctacccacaccatgccacacagctccatgcccaccagccgcagccggctaccacgcctactggaagccag ccgcagtcccagcatgcggcccccagtcctgtccaggtgcctgccatggggggtgctgagtggtcctggtgcaggaatgg gtggccagaagaagggatagagctaggggtcatttctgagtggcgaggactgggggccagcgagttgctggcctgtgtgg cactcaaccttcccctccccagcagcatcaggcggggcaggccccacacttgggcagtggacagccacagcagaatctgt accacccaggggccctgacaggcacgccgcccrctctgccaccgggaccttctgcccagtcccctcagagcagcrtcccc cagccagccgctgtgtargccarccaccaccagcagctgccocacggcttcaccaacatggcccatgttacccaggccca tgtccaaactggaatcacagcagccccgccccctcaccctggggctccccacccgccccaggtgatgctgctgcacccac cccagagtcatggggggcccccccaaggcgcggtgccccagagtggggtgcctgcactctcagcttccacaccctcaccc tacccatacatcggacacccccaaggtgagcagcctggccaggcgcctggatttccaggaggagccgatgacaggattcg tgagtτctcattagctgggggaa ttggcatggaagagctgaggggatgcaggtggggcagggatgacacgggttctggg tggggagtgaggggtcttggaggcagggctgtcccacagggcgcccgctgacctgcacctgtctgtgaagtatgtagggt gggcagaagccacagtcgccgccσccaggggcttgctcctggccctgtcct tgcrtccctccgtcctcgctcagttgtg a ccagcagcσcccctccccactαcctccccaαtctcaαtqaccccαatαtctctσacttaσcggaggtaagσtcaqτqc agcagacagggccagactggggtgtggggggctgagctgggcacatgagtgagggtrctggcttacigggaaacagcgat tgacctgtgcttctgaoagcccccgagacactrgaggagccgctctttcccagacacacccccacgcccccactggacgg cattggaggaaggacagctgcttgggttctaatgctcctgctctcttctctttcccctccaaccagttcaatctcatccc tcccagcagctccccttccaccccccggggaactgaagattgtcctggccgcgaccrgagacctccatgagtggagggaa gagtgatctatgtctcttcccccagcagctcggaccactcccagccccccatccccccgrtccccaggggagctggggaa ttcctgccaagcaccttgaatgggaggggcctcacagagggcagggccagggtccagcaggggtggggggttcctgttct gccccrgcccgtccccacccagtcttgccctcccatcctctcatc attcccccgcrggagacggaagaccttttatttt ctatratttataacttcagacttgggccccc gttctttctttcccattaacttgagtgacctgtgtgagagacagacag atgccccacgaggatggctggacaaggacttt actttttattacataaaaatat'-aaaaaaεaaataaaaaaaataaaa ttttaaacraacttaaaaaaaaaaaaaaaaaaaa

' Anucleoude sequence oi a human alaxιn-2 like gene (SEQ ID NO. 1).

Tabk2^b

MGWRPGRQAPHFPPAARPPPLPPRGAΞSRRGFLSPPPTPRGSPRPPTAGPGCPIPLAΞRALQRGPSPGP SPSLLSNSPS

GRCHHVEASAATTALPAPAAAPHATCRGPSAPRGHSPPNGGLPGPIΛTSAAPPGPPAAASPCLGPVAAAGSGLRRGAEGI

LAPQPPPPQQHQERPGAAAIGSARGQSTGKGPPQSPVFEGVYNNSRMLHF TAWGΞTCDVKVKNGTTYEGIFKTLSSKF

ELAVDAVHRKASEPAGGPRREDIVDTMVFKPSDVMLVHFRNVDFNYATKDKF QSAIAMNSKVNGEHKEKVLQRWEGGDΞ

NSDDYD ESD SNGWDPNEMFKFNEENYGVKTTYDΞS ΞSYTVPLE DNSEEFRQRELRAAQ AREIESSPQYRLRIAME

NDDGRTEEEKHSAVQRQGSGRESPS1ASREGKYIPLPQRVREGPRGGVRCΞSΞRGGRPGLΞSLPPRGPHHLDNSSPGPGS

EARGINGGPSR SPKAQRPLRGAKT SSPSNR LENFCSTSSCSGPDVSPRSPKSAAPAPISASCPEPPIGSAVPTSSAS

IPV SSVSDPGVGSISPASPKIS APTDV ELSTKEPGRT EPQELARIAG VPGLQNEQKRFQLEELR FGAQF QPS

SSPENSLDPFPPRI KEEPKG EKEVDG TΞEPMGSPVSSKTESVSDKEDKPPLAPSGGTEGPEQPPPPCPΞQTGSPPV

GLIKGEDKDEGPVAEQVKKΞTLNPNAKEFNPTKPL ΞVNKSTΞTPTSPGPRTHSTPSIPV TAGQSGLYSPQYIΞYIPQI

HMGPAVQAPQMYPYPVSNSVPGQQGKYRGAKGSLPPQRSDQHQPASAPPMMQAAAAAGPPLVAATPYSSYIPYNPQQFPG

QPAM QPMAHYPSQPVFAPMLQSNPRMLTSGSHPQAIVSΞΞTPQYPSAEQPTPQA YATVHQΞYPHHATQLHAHQPQPAT

TPTGSQPQΞQHAAPΞPVQVPAMGGAEWSWCRNGWPEEGIELGVIΞEWRGLGASE LACVALNLP PΞΞIRRGRPHT AVD SHSRICTTQGP An ammo acid sequence of a human ataxιn-2 like gene (SEQ ID NO: 2).

One polynucleotide of the present mvention encoding the ataxιn-2 like gene may be obtained using standard cloning and screening, from a cDNA library derived from mRNA in cells of human adult brain using the expressed sequence tag (EST) analysis (Adams, M.D., et al. Science (1991) 252:1651-1656; Adams, M.D. et ai., Nature, (1992) 555:632-634; Adams, M.D., et ai, Nature (1995) 377 Supp.3-174). Polynucleotides of the mvention can also be obtained from natural sources such as genomic DNA libraries or can be synthesized usmg well known and commercially available techniques.

The nucleotide sequence encoding ataxin-2 like gene polypeptide of SEQ ID NO:2 may be identical to the polypeptide encoding sequence contained in Table 1 (nucleotide number 63 to 3215 of SEQ ID NO: 1), or it may be a sequence, which as a result of the redundancy (degeneracy) of the genetic code, also encodes the polypeptide of SEQ ID NO:2. When the polynucleotides of the invention are used for the recombinant production of the ataxin-2 like gene polypeptide, the polynucleotide may include the coding sequence for the mature polypeptide or a fragment thereof, by itself; 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. For example, a marker sequence which facilitates purification of the fused polypeptide can be encoded. In certain preferred embodiments of this aspect of the invention, the marker sequence is a hexa-histidine peptide, as provided in the pQE vector (Qiagen, Inc.) and described in Gentz et al. , Proc Natl Acad Sci 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, ribosome binding sites and sequences that stabilize mRNA.

Further preferred embodiments are polynucleotides encoding ataxin-2 like gene variants which comprise the amino acid sequence ataxin-2 like gene polypeptide of Table 2 (SEQ ID NO:2) in which several, 5-10, 1-5, 1-3, 1-2 or 1 amino acid residues are substituted, deleted or added, in any combination.

The present invention further relates to polynucleotides that hybridize to the herein above- described sequences. In this regard, the present invention especially relates to polynucleotides which hybridize under stringent conditions to the herein above-described polynucleotides. As herein used, the term "stringent conditions" means hybridization 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 in SEQ ID NO: 1 or a fragment thereof, may be used as hybridization probes for cDNA and genomic DNA, to isolate fiill-length cDNAs and genomic clones encoding ataxin-2 like gene polypeptides, 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 similarity to the ataxin-2 like gene. Such hybridization techniques are known to those of skill in the art. Typically these nucleotide sequences are 80% identical, preferably 90% identical, more preferably 95% identical to that of the referent. The probes generally will comprise 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.

In one embodiment, to obtain a polynucleotide encoding the ataxin-2 like gene polypeptide, including homologs and orthologs from species other than human, a method comprises the steps of screening an appropriate library under stingent hybridization conditions with a labeled probe having the SEQ ID NO 1 or a fragment thereof, and isolating full-length cDNA and genomic clones containing said polynucleotide sequence Thus in another aspect, ataxιn-2 like gene polynucleotides of the present invention further mclude a nucleotide sequence comprising a nucleotide sequence that hybπdize under stringent condition to a nucleotide sequence having SEQ ED NO 1 or a fragment thereof Also mcluded with atax -2 like gene polypeptides are polypeptides comprising ammo acid sequences encoded by nucleotide sequences obtained by the above hybπdization condition Such hybndization techniques are well known to those of skill m the art Stringent hybndization conditions are as defined above or, alternatively, conditions under overnight incubation at 42°C m a solution compπsmg 50% formamide, 5xSSC (150mM NaCl, 15mM tπsodium citrate), 50 mM sodium phosphate (pH7 6), 5x Denhardt-s solution, 10 % dextran sulfete, and 20 microgram/ml denatured, sheared salmon sperm DNA, followed by washing the filters in 0 lx SSC at about 65°C

The polynucleotides and polypeptides of the present mvention may be employed as research reagents and materials for discovery of treatments and diagnostics to animal and human disease

Vectors, Host Cells, Expression

The present mvention also relates to vectors which comprise a polynucleotide or polynucleotides of the present mvention, and host cells which are genetically engineered with vectors of the mvention and to the production of polypeptides of the mvention by recombinant techniques Cell-free translation systems can also be employed to produce such proteins using RNAs deπved from the DNA constructs of the present mvention

For recombinant production, 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 descπbed in many standard laboratory manuals, such as Davis et ai, BASIC METHODS M MOLECULAR BIOLOGY (1986) and Sambrook et al , MOLECULAR CLONING A LABORATORY MANUAL, 2nd Ed , Cold Spπng Harbor Laboratory Press, Cold Spπng Harbor, NY ( 1989) such as calcium phosphate transfection, DEAE-dextran mediated transfection, transvection, microinjection, canonic lipid-mediated transfection, electroporation, transduction, scrape loading, ballistic introduction or infection

Representative examples of appropπate hosts mclude bacteπal cells, such as streptococci, staphylococci, E coh, Streptomyces and Bacillus subtihs cells, fungal cells, such as yeast cells and Aspergillus cells, insect cells such as Drosophila S2 and Spodoptera Sf9 cells, animal cells such as CHO, COS, HeLa, C127, 3T3, BHK. HEK 293 and Bowes melanoma cells, and plant cells

A great vaπety of expression systems can be used Such systems mclude, among others, chromosomal, episomal and virus-deπved systems, e g , vectors deπved from bacteπal plasmids, from bacteπophage, 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, fowl pox viruses, pseudorabies viruses and retrovuuses, and vectors deπved from combinations thereof, such as those deπved from plasmid and bacteπophage genetic elements, such as cosmids and phagemids The expression systems may contam control regions that regulate as well as engender expression Generally, any system or vector suitable to maintain, propagate or express polynucleotides to produce a polypeptide in a host may be used The appropπate nucleotide sequence may be inserted into an expression system by any of a vaπety of well-known and routine techniques, such as, for example, those set forth Sa brook et al , MOLECULAR CLONING, A LABORATORY MANUAL (supra)

For secretion of the translated protem mto the lumen of the endoplasmic reticulum, mto the peπplasrmc space or into the extracellular environment, appropπate secretion signals may be incorporated mto the desired polypeptide These signals may be endogenous to the polypeptide or they may be heterologous signals. If the ataxιn-2 like gene polypeptide is to be expressed for use in screening assays, generally, it is preferred that the polypeptide be produced at the surface of the cell In this event, the cells may be harvested pπor to use m the screenmg assay If the ataxιn-2 like gene polypeptide is secreted mto the medium, the medium can be recovered m order to recover and puπfy the polypeptide; if produced intracellularly, the cells must first be lysed before the polypeptide is recovered Ataxιn-2 like gene polypeptides can be recovered and purified from recombinant cell cultures by well-known methods mcludmg ammonium sulfete or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography and lectin chromatography Most preferably, high performance liquid chromatography is employed for puπfication Well known techniques for refolding proteins may be employed to regenerate active conformation when the polypeptide is denatured during isolation and or purification

Diagnostic Assays

This mvention also relates to the use of ataxιn-2 like gene polynucleotides for use as diagnostic reagents Detection of a mutated form of the ataxιn-2 like 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 ataxιn-2 like gene Individuals carrying mutations the ataxιn-2 like gene may be detected at the DNA level by a vaπety of techniques Nucleic acids for diagnosis may be obtained from a subject's cells, such as from blood, uπne, saliva, tissue biopsy or autopsy mateπal. The genomic DNA may be used directly for detection or may be amplified enzymatically by using PCR or other amplification techniques pnor to analysis. RNA or cDNA may also be used in similar fashion. Deletions and insertions can be detected by a change m size of the amplified product in comparison to the normal genotype. Point mutations can be identified by hybridizing amplified DNA to labeled ataxin-2 like gene nucleotide sequences. Perfectly matched sequences can be distinguished from mismatched duplexes by RNase digestion or by differences m melting temperatures. DNA sequence differences may also be detected by alterations m electrophoretic mobility of DNA fragments in gels, with or without denaturing 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 et al., Proc NatlAcadSci USA (1985) 85: 4397^401. In another embodiment, an array of oligonucleotide probes compπsmg ataxιn-2 like gene nucleotide sequences or fragments thereof can be constructed to conduct efficient screening of e.g., genetic mutations. Array technology methods are well known and have general applicability and can be used to address a vaπety of questions in molecular genetics including gene expression, genetic linkage, and genetic variability. (See for example: M.Chee etai., Science, Vol 274, pp 610-613 (1996)).

The diagnostic assays offer a process for diagnosing or determining a susceptibility to ataxia, cardiomyopathy, Deafness, neurological disease, cancer, and AIDS through detection of mutation in the ataxin-2 like gene by the methods described.

In addition, ataxia, cardiomyopathy, Deafness, neurological disease, cancer, and AIDS can be diagnosed by methods compπsmg determining from a sample derived from a subject an abnormally decreased or increased level of ataxιn-2 like gene polypeptide or ataxιn-2 like gene mRNA Decreased or increased 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 hybridization methods. Assay techniques that can be used to determine levels of a protein, such as an ataxin-2 like gene polypeptide, in a sample denved from a host are well-known to those of skill in the art. Such assay methods mclude radioimmunoassays, competitive-binding assays, Western Blot analysis and ELISA assays. Thus m another aspect, the present mvention relates to a diagonostic kit for a disease or suspectability to a disease, particularly ataxia, cardiomyopathy, deafness, neurological disease, cancer, and AIDS, which compπses:

(a) an ataxin-2 like gene polynucleotide, preferably the nucleotide sequence of SEQ ID NO 1, or a fragment thereof ; (b) a nucleotide sequence complementary to that of (a),

(c) an ataxιn-2 like gene polypeptide, preferablv the polypeptide of SEQ ID NO 2, or a fragment thereof, or

(d) an antibody to an ataxιn-2 like gene polypeptide, preferably to the polypeptide of SEQ ID NO 2

It will be appreciated that m any such kit, (a), (b), (c) or (d) may compπse a substantial component

Chromosome Assays The nucleotide sequences of the present mvention are also valuable for chromosome identification The sequence is specifically targeted to and can hybπdize with a particular location on an individual human chromosome The mapping of relevant sequences to chromosomes according to the present mvention is an important first step m correlating those sequences with gene associated disease Once a sequence has been mapped to a precise chromosomal location, the physical position of the sequence on the chromosome can be correlated with genetic map data Such data are found, for example, in V McKusick, Mendehan Inheπtance 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 (coinheπtance 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 m some or all of the affected individuals but not m any normal individuals, then the mutation is likely to be the causative agent of the disease

The human ataxιn-2 like gene was mapped to chromosome 9cen where familial dilated cardiomyopathy, deafness, cartilage-hair hypoplasia, Melkersson-Rosenthal syndrome, and inclusion body myopathy, among others, were localized

Antibodies

The polypeptides of the mvention or their fragments or analogs thereof, or cells expressing them can also be used as lmmunogens to produce antibodies immunospecific for the ataxιn-2 like gene polypeptides The teim "immunospecific" means that the antibodies have substantial! greater affinity for the polypeptides of the mvention than their affinity for other related polypeptides m the pπor art Antibodies generated against the ataxιn-2 like gene polypeptides can be obtained by administering me polypeptides or epitope-beaπng fragments, analogs or cells to an animal, preferably a nonhuman, usmg routine protocols For preparation of monoclonal antibodies, any technique which provides antibodies produced by continuous cell line cultures can be used Examples mclude the hybndoma technique (Kohler, G and Milstein, C , Nature (1975) 256495-497), the tπoma technique, the human B-cell hybndoma technique (Kozbor et al , Immunology Today (1983) 4 72) and the EBV- hybπdoma technique (Cole et al , MONOCLONAL ANTIBODIES AND CANCER THERAPY, pp. 77-96, Alan R Liss, Inc , 1985)

Techniques for the production of single cham antibodies (U S Patent No 4,946,778) can also be adapted to produce single cham antibodies to polypeptides of this mvention Also, transgemc mice, or other organisms mcludmg other mammals, may be used to express humanized antibodies

The above-descπbed antibodies may be employed to isolate or to identify clones expressing the polypeptide or to puπfy the polypeptides bv affinity chromatography

Antibodies against ataxιn-2 like gene polypeptides may also be employed to treat ataxia, cardiomyopathy, deafness, neurological disease, cancer, and AIDS, among others

Vaccines

Another aspect of the mvention relates to a method for mducmg an immunological response m a mammal which compπses inoculating the mammal with ataxιn-2 like gene polypeptides, or fragments thereof, adequate to produce antibody and/or T cell immune response to protect said animal from ataxia, cardiomyopathy, deafness, neurological disease, cancer, and AIDS, among others Yet another aspect of the mvention relates to a method of mducmg immunological response m a mammal which compπses delivering ataxm-2 like gene polypeptide via a vector directing expression of ataxιn-2 like gene polynucleotide m vivo in order to mduce such an immunological response to produce antibody to protect said animal from diseases

A further aspect of the mvention relates to an lmmunological/vaccine formulation (composition) which, when introduced mto a mammalian host, mduces an immunological response m that mammal to an ataxιn-2 like gene polypeptide wherein the composition compπses an ataxιn-2 like gene polypeptide or ataxιn-2 like gene The vaccme formulation may further compπse a suitable earner Since ataxιn-2 like gene polypeptides may be broken down m the stomach, it is preferably administered parenterally (mcludmg subcutaneous, intramuscular, intravenous, lntradermal etc injection) Formulations suitable for parenteral administration mclude aqueous and non-aqueous steπle injection solutions which may contain anti-oxidants, buffers, bacteπostats 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 example, sealed ampoules and vials and may be stored in a freeze-dried condition requiring only the addition of the sterile liquid carrier immediately prior to use. The vaccine formulation may also include adjuvant systems for enhancing the immunogenicity of the formulation, such as oil-in water systems and other systems known in the art. The dosage will depend on the specific activity of the vaccine and can be readily determined by routine experimentation.

Screening Assays The ataxin-2 like gene polypeptides 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 ataxin-2 like gene polypeptide of the present invention. Thus, polypeptides of the invention may also be used to assess identify agonist or antagonists from, for example, cells, cell-free preparations, chemical libraries, and natural product mixtures. These 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 structural or functional mimetics of the polypeptide of the present invention. See Coligan et al, Current Protocols in Immunology l(2):Chapter 5 (1991).

Ataxin-2 like gene polypeptides are responsible for many biological functions, including many pathologies. Accordingly, it is desirous to find compounds and drugs which stimulate ataxin-2 like gene polypeptides on the one hand and which can inhibit the function of ataxin-2 like gene polypeptides on the other hand. In general, agonists are employed for therapeutic and prophylactic purposes for such conditions as ataxia, cardiomyopathy, deafness, neurological disease, cancer, and AIDS. Antagonists may be employed for a variety of therapeutic and prophylactic purposes for such conditions as ataxia, cardiomyopathy, deafness, neurological disease, cancer, and AIDS. In general, such screening procedures may involve using appropriate cells which express the ataxin-2 like gene polypeptide or respond to the ataxin-2 like gene polypeptide of the present invention. Such cells include cells from mammals, yeast, Drosophila or E. coli. Cells which express the ataxin-2 like gene polypeptide (or cell membrane containing the expressed polypeptide) or respond to the ataxin-2 like gene polypeptide are then contacted with a test compound to observe binding, 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 ataxin-2 like gene activity. The assays may simply test binding of a candidate compound wherein adherence to the cells bearing the ataxin-2 like gene 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 m a signal generated by activation of the ataxιn-2 like gene polypeptide, usmg detection systems appropπate to the cells beaπng the ataxιn-2 like gene polypeptide Inhibitors of activation are generally assayed m the presence of a known agonist and the effect on activation by the agonist by the presence of the candidate compound is observed

Further, the assays may simply comprise the steps of mixing a candidate compound with a solution containing an ataxιn-2 like gene polypeptide to form a mixture, measuring ataxιn-2 like gene activity in the mixture, and comparing the ataxιn-2 like gene activity of the mixture to a standard The ataxm-2 like gene cDNA, protem and antibodies to the protem may also be used to configure assays for detectmg the effect of added compounds on the production of ataxιn-2 like gene mRNA and protem m cells. For example, an ELISA may be constructed for measuring secreted or cell associated levels of ataxιn-2 like gene protem usmg 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 atax -2 like gene (also called antagonist or agomst, respectively) from suitably manipulated cells or tissues.

The ataxιn-2 like gene protem may be used to identify membrane bound or soluble receptors, if any, through standard receptor binding techniques known m the art These mclude, but are not limited to, ligand binding and cross nking assays in which the ataxιn-2 like gene is labeled with a radioactive isotope (eg 1251), chemically modified (eg biotinylated), or fused to a peptide sequence suitable for detection or purification, and mcubated with a source of the putative receptor (cells, cell membranes, cell supernatants, tissue extracts, bodily fluids) Other methods mclude biophysical techniques such as surface plasmon resonance and spectroscopy In addition to bemg used for puπfication and cloning of the receptor, these binding assays can be used to identify agonists and antagonists of the ataxιn-2 like gene which compete with the binding of the ataxιn-2 like gene to its receptors, if any Standard methods for conducting screenmg assays are well understood m the art

Examples of potential ataxιn-2 like gene polypeptide antagonists mclude antibodies or, in some cases, oligonucleotides or proteins which are closely related to the ligands, substrates, receptors, enzymes, etc., as the case may be, of the ataxιn-2 like gene polypeptide, e g , a fragment of the ligands, substrates, receptors, enzymes, etc , or small molecules which bmd to the polypetide of the present mvention but do not elicit a response, so that the activity of the polypeptide is prevented

Thus m another aspect, the present mvention relates to a screenmg kit for identifying agonists, antagonists, ligands, receptors, substrates, enzymes, etc for ataxιn-2 like gene polypeptides, or compounds which decrease or enhance the production of ataxιn-2 like gene polypeptides, which compπses

(a) an ataxm-2 like gene polypeptide, preferably that of SEQ ID NO 2,

(b) a recombinant cell expressmg an ataxιn-2 like gene polypeptide, preferably that of SEQ ID NO 2,

(c) a cell membrane expressmg an ataxιn-2 like gene polypeptide, preferably that of SEQ ID NO 2, or

(d) antibody to an ataxιn-2 like gene polypeptide, preferably that of SEQ ID NO 2

It will be appreciated that m any such kit, (a), (b), (c) or (d) may compπse a substantial component

Prophylactic and Therapeutic Methods

This mvention provides methods of treating abnormal conditions such as ataxia, cardiomyopathy, deafness, neurological disease, cancer, and AIDS, related to both an excess of and insufficient amounts of, ataxm-2 like gene polypeptide activity

If the activity of the ataxιn-2 like gene polypeptide is in excess, several approaches are available One approach compπses administering to a subject an inhibitor compound (antagonist) as hereinabove descnbed along with a pharmaceutically acceptable earner in an amount effective to inhibit the function of the ataxιn-2 like gene polypeptide. such as, for example, by blockmg 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 ataxιn-2 like gene polypeptides still capable of binding the ligand, substrate, enzymes, receptors, etc m competition with endogenous ataxιn-2 like gene polypeptides may be administered Typical embodiments of such competitors compπse fragments of the ataxm-2 like gene polypeptide In still another approach, expression of the gene encoding endogenous ataxιn-2 like gene polypeptide can be inhibited usmg expression blockmg techmques Known such techmques mvolve the use of antisense sequences, either internally generated or separately administered See, for example, O'Connor, JNeurochem (1991) 56 560 in Oligodeoxynucleotides as Antisense Inhibitors of Gene Expression. CRC Press, Boca Raton, FL (1988) Alternatively, oligonucleotides which form tπple helices with the gene can be supphed See, for example, Lee et al , Nucleic Acids Res (1979) 6 3073, Cooney et ai , Science (1988) 241 456, Dervan et al , Science (1991) 251 1360 These ohgomers can be administered per se or the relevant oligomers can be expressed m vivo For treating abnormal conditions related to an under-expression of ataxιn-2 like gene and its activity, several approaches are also available One approach compπses administering to a subject a therapeutically effective amount of a compound which activates the ataxιn-2 like gene polypeptide, i e , an agonist as descπbed above, in combination with a pharmaceutically acceptable earner, to thereby alleviate the abnormal condition Alternatively, gene therapy may be employed to effect the endogenous production of ataxιn-2 like gene by the relevant cells the subject For example, a polynucleotide of the mvention may be engmeered for expression m a rephcation defective retroviral vector, as discussed above The retroviral expression construct may then be isolated and introduced mto a packaging cell transduced with a retroviral plasmid vector containing RNA encoding a polypeptide of the present invention such that the packaging 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 expression of the polypeptide m vivo For overview of gene therapy, see Chapter 20, Gene Therapy and other Molecular Genetic-based Therapeutic Approaches, (and references cited therein) in Human Molecular Genetics, T Strachan and A P Read, BIOS Scientific Publishers Ltd (1996) Another approach is to administer a therapeutic amount of ataxιn-2 like gene polypeptides m combination with a suitable pharmaceutical earner

Formulation and Administration

Peptides, such as the soluble form of ataxιn-2 like gene polypeptides, and agonists and antagonist peptides or small molecules, may be formulated in combination with a suitable pharmaceutical earner. Such formulations compπse a therapeutically effective amount of the polypeptide or compound, and a pharmaceutically acceptable earner or excipient. Such earners mclude 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 compnsmg 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 mclude injection, typically by intravenous injection Other injection routes, such as subcutaneous, intramuscular, or lntrapeπtoneal, can be used Alternative means for systemic administration mclude transmucosal and transdermal administration usmg penetrants such as bile salts or fusidic acids or other detergents In addition, if properly formulated entenc or encapsulated formulations, 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 The dosage range required depends on the choice of peptide, the route of administration, the nature of the formulation, the nature of the subject's condition, and the judgment of the attending practitioner. Suitable dosages, however, are in me range of 0.1 - 100 μg/kg of subject. Wide variations in the needed dosage, however, are to be expected in view of the variety of compounds available and the differing efficiencies of various routes of administration. For example, oral administration would be expected to require higher dosages than administration by intravenous injection. Variations in these dosage levels can be adjusted using standard empirical routines for optimization, as is well understood in the art.

Polypeptides used in treatment can also be generated endogenously in the subject, in treatment modalities often referred to as "gene therapy" as described above. Thus, for example, 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 plasmid vector. The cells are then introduced into the subject.

All publications, including but not limited to patents and patent applications, cited in this specification are herein incorporated by reference as if each individual publication were specifically and individually indicated to be incorporated by reference herein as though fully set forth.

SEQUENCE LISTING

(1) GENERAL INFORMATION

(i) APPLICANT: Hunan Medical University

(ii) TITLE OF THE INVENTION: An Ataxin-2 Like Gene

(iii) NUMBER OF SEQUENCES: 2

(iv) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: RATNER & PRESTIA

(B) STREET: P.O. BOX 980

(C) CITY: VALLEY FORGE

(D) STATE: PA

(E) COUNTRY: USA (F) ZIP: 19482

(V) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Diskette (B) COMPUTER: IBM Compatible

(C) OPERATING SYSTEM: DOS

(D) SOFTWARE: FastSEQ for Windows Version 2.0

(vi) CURRENT APPLICATION DATA:

(A) APPLICATION NUMBER: TO BE ASSIGNED

(B) FILING DATE:

(C) CLASSIFICATION: UNKNOWN

(vii) PRIOR APPLICATION DATA:

(A) APPLICATION NUMBER:

(B) FILING DATE:

(viii) ATTORNEY/AGENT INFORMATION:

(A) NAME: PRESTIA, PAUL F

(B) REGISTRATION NUMBER: 23,031

(C) REFERENCE/DOCKET NUMBER: GH-70348

(ix) TELECOMMUNICATION INFORMATION:

(A) TELEPHONE: 610-407-0700

(B) TELEFAX: 610-407-0701

(C) TELEX: 846169

(2) INFORMATION FOR SEQ ID NO:lι

(i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 4674 base pairs

(B) TYPE: nucleic acid

(C) STRANDEDNESS : single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: cDNA

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:l:

CGGGACGCTC CTCACTTCCT AGATGGGATG GCGGCCGGGA AGAGGCGCTC CTCATTTCCT 60 AGATGGGATG GCGGCCGGGC AGAGATGCTC CTCACTTTCC TCCAGCCGCG AGACCCCCTC 120

CCCTTCCGCC TCGCGGCGCT TCCTCGCGCC GCGGTTTTCT CTCTCCACCC CCGACACCGC 180

GGGGCTCCCC CCGCCCGCCC ACGGCGGGCC CCGGCTGCCC GATCCCCCTC GCTTCCCGCG 240

CTCTCCAGCG GGGCCCCAGC CCCGGCCCCC TCTCTCCCTC CCTTCTCTCT AATTCCCCTT 300 CCGGACGCTG CCATCATGTT GAAGCCTCAG CCGCTACAAC AGCCCTCCCA GCCCCAGCAG 360 CCGCCCCCCA CGCAACATGC CGTGGCCCGT CGGCCCCCCG GGGGCACAGC CCTCCCAACG 420 GCGGCCTCCC GGGGCCGCTG GCCACCTCTG CGGCTCCTCC CGGGCCTCCA GCGGCCGCCT 480 CCCCCTGCCT GGGGCCTGTG GCCGCTGCCG GGAGCGGGCT CCGCCGGGGA GCCGAAGGCA 540 TCTTGGCGCC GCAGCCGCCG CCGCCGCAGC AACACCAGGA GAGGCCGGGG GCAGCCGCCA 600 TCGGCAGCGC CAGGGGACAG AGCACAGGAA AGGGACCCCC ACAGTCACCT GTGTTTGAAG 660 GCGTCTACAA CAATTCCAGA ATGCTGCATT TCCTTACAGC TGTTGTGGGC TCCACTTGTG 720 ATGTAAAGGT GAAAAATGGT ACCACTTATG AGGGTATCTT CAAGACGCTA AGCTCAAAGT 780 TTGAACTAGC CGTGGATGCT GTGCACCGGA AAGCATCTGA GCCAGCAGGT GGCCCTCGTC 840 GGGAGGACAT TGTGGACACC ATGGTGTTTA AGCCAAGTGA TGTCATGCTT GTTCACTTCC 900 GAAATGTTGA CTTCAACTAT GCTACTAAAG ACAAGTTCAC CGATTCAGCC ATTGCCATGA 960

ACTCGAAAGT GAATGGGGAA CACAAAGAGA AGGTGCTTCA GCGCTGGGAG GGGGGTGACA 1020

GCAACAGCGA CGACTATGAC CTCGAGTCTG ACATGTCCAA TGGATGGGAC CCCAATGAAA 1080

TGTTCAAGTT CAATGAGGAG AACTACGGTG TGAAGACTAC CTATGATAGC AGTCTTTCTT 1140 CTTATACGGT GCCCTTAGAA AAGGACAACT CAGAAGAGTT TCGTCAGCGA GAGCTGCGTG 1200

CGGCCCAGTT GGCTCGAGAG ATTGAATCAA GCCCCCAGTA CCGCCTACGG ATCGCCATGG 1260

AGAACGACGA TGGGCGCACT GAAGAGGAGA AGCACAGTGC AGTCCAGCGG CAGGGCTCAG 1320

GGCGGGAGAG CCCCAGCTTG GCATCCAGGG AGGGGAAGTA TATCCCTCTG CCTCAACGAG 1380

TCCGGGAAGG TCCCCGGGGA GGAGTTCGAT GCAGCAGCTC TCGGGGCGGT CGGCCTGGCC 1440 TTAGCTCTTT GCCACCTCGT GGCCCTCACC ATCTGGACAA CAGCAGCCCT GGCCCAGGTT 1500

CTGAGGCCCG TGGTATCAAT GGAGGCCCTT CCCGCATGTC CCCAAAGGCA CAGCGGCCTC 1560

TGAGAGGTGC CAAGACTCTG TCTTCGCCCA GTAATAGGCT TCTGGAGAAC TTCTGTTCCA 1620

CCTCCTCCTG CAGTGGGCCG GATGTATCCC CGCGTTCTCC CAAGTCTGCT GCCCCTGCCC 1680

CAATCTCAGC TTCCTGTCCA GAGCCTCCCA TCGGCTCGGC AGTGCCAACC TCTTCAGCCT 1740 CCATCCCTGT GACCTCATCA GTCTCAGATC CTGGAGTGGG CTCCATTTCT CCAGCTTCTC 1800

CAAAGATCTC CCTGGCCCCC ACAGATGTAA AAGAACTCTC TACCAAGGAA CCTGGGAGAA 1860

CTCTGGAGCC CCAGGAGCTG GCTCGGATAG CTGGGAAAGT CCCTGGTCTT CAGAATGAAC 1920

AGAAACGATT CCAACTGGAA GAACTGAGAA AGTTTGGGGC CCAGTTTAAG CTTCAGCCCA 1980

GTAGCTCCCC TGAGAACAGC CTGGATCCTT TTCCTCCCCG GATCTTAAAG GAGGAGCCCA 2040 AAGGAAAGGA GAAAGAGGTT GATGGTCTGT TGACTTCAGA GCCCATGGGG TCTCCCGTCT 2100

CCTCCAAGAC AGAGTCCGTA TCGGATAAGG AGGACAAACC ACCCCTGGCA CCATCAGGAG 2160

GCACTGAGGG GCCAGAGCAG CCCCCACCAC CTTGTCCAAG CCAAACTGGC AGCCCCCCGG 2220

TGGGCCTCAT CAAGGGAGAA GACAAAGATG AGGGCCCTGT TGCTGAACAA GTAAAGAAAT 2280

CAACGTTGAA CCCTAATGCT AAGGAGTTCA ATCCTACAAA GCCTCTGCTG TCTGTGAATA 2340 AATCCACCAG TACCCCAACT TCTCCGGGGC CCCGGACTCA TTCAACTCCC TCCATCCCGG 2400

TGCTGACAGC AGGCCAGAGT GGGCTATACA GCCCCCAGTA CATCTCCTAC ATACCTCAGA 2460

TCCACATGGG ACCAGCTGTG CAGGCACCTC AGATGTATCC ATATCCTGTA TCCAATTCAG 2520

TGCCTGGGCA GCAGGGCAAG TACCGGGGAG CAAAAGGCTC CCTTCCTCCG CAGCGCTCGG 2580

ACCAACACCA GCCAGCCTCA GCCCCGCCGA TGATGCAGGC CGCCGCGGCT GCTGGCCCGC 2640 CTCTGGTGGC TGCCACGCCC TATTCTTCCT ACATCCCCTA CAACCCTCAG CAGTTCCCAG 2700

GCCAGCCAGC CATGATGCAG CCCATGGCCC ACTACCCCTC ACAGCCGGTG TTTGCCCCCA 2760

TGCTTCAGAG CAACCCACGC ATGCTGACGT CGGGCAGCCA TCCCCAGGCC ATCGTGTCAT 2820

CCTCTACCCC TCAATACCCT TCTGCAGAGC AGCCTACCCC CCAAGCCCTT TATGCCACTG 2880 TTCACCAGTC CTACCCACAC CATGCCACAC AGCTCCATGC CCACCAGCCG CAGCCGGCTA 2940

CCACGCCTAC TGGAAGCCAG CCGCAGTCCC AGCATGCGGC CCCCAGTCCT GTCCAGGTGC 3000

CTGCCATGGG GGGTGCTGAG TGGTCCTGGT GCAGGAATGG GTGGCCAGAA GAAGGGATAG 3060

AGCTAGGGGT CATTTCTGAG TGGCGAGGAC TGGGGGCCAG CGAGTTGCTG GCCTGTGTGG 3120

CACTCAACCT TCCCCTCCCC AGCAGCATCA GGCGGGGCAG GCCCCACACT TGGGCAGTGG 3180

ACAGCCACAG CAGAATCTGT ACCACCCAGG GGCCCTGACA GGCACGCCGC CCTCTCTGCC 3240

ACCGGGACCT TCTGCCCAGT CCCCTCAGAG CAGCTTCCCC CAGCCAGCCG CTGTGTATGC 3300

CATCCACCAC CAGCAGCTGC CCCACGGCTT CACCAACATG GCCCATGTTA CCCAGGCCCA 3360

TGTCCAAACT GGAATCACAG CAGCCCCGCC CCCTCACCCT GGGGCTCCCC ACCCGCCCCA 3420

GGTGATGCTG CTGCACCCAC CCCAGAGTCA TGGGGGGCCC CCCCAAGGCG CGGTGCCCCA 3480

GAGTGGGGTG CCTGCACTCT CAGCTTCCAC ACCCTCACCC TACCCATACA TCGGACACCC 3540

CCAAGGTGAG CAGCCTGGCC AGGCGCCTGG ATTTCCAGGA GGAGCCGATG ACAGGATTCG 3600

TGAGTTCTCA TTAGCTGGGG GAATTTGGCA TGGAAGAGCT GAGGGGATGC AGGTGGGGCA 3660

GGGATGACAC GGGTTCTGGG TGGGGAGTGA GGGGTCTTGG AGGCAGGGCT GTCCCACAGG 3720

GCGCCCGCTG ACCTGCACCT GTCTGTGAAG TATGTAGGGT GGGCAGAAGC CACAGTCGCC 3780

GCCGCCAGGG GCTTGCTCCT GGCTCTGTCC TTTGCTTCCC TCCGTCCTCG CTCAGTTGTG 3840

ATCCAGCAGC CCCCCTCCCC ACTGCCTCCC CAGTCTCAGT GACCCCGATG TCTCTGACTT 3900

AGCGGAGGTA AGGTCAGTGC AGCAGACAGG GCCAGACTGG GGTGTGGGGG GCTGAGCTGG 3960

GCACATGAGT GAGGGTTCTG GCTTACTGGG AAACAGCGAT TGACCTGTGC TTCTGACAGC 4020

CCCCGAGACA CTTGAGGAGC CGCTCTTTCC CAGACACACC CCCACGCCCC CACTGGACGG 4080

CATTGGAGGA AGGACAGCTG CTTGGGTTCT AATGCTCCTG CTCTCTTCTC TTTCCCCTCC 4140

AACCAGTTCA ATCTCATCCC TCCCAGCAGC TCCCTTTCCA CCCCCCGGGG AACTGAAGAT 4200

TGTCCTGGCC GCGACCTGAG ACCTCCATGA GTGGAGGGAA GAGTGATCTA TGTCTCTTCC 4260

CCCAGCAGCT CGGACCACTC CCAGCCCCCC ATCCCCCCGT TCCCCAGGGG AGCTGGGGAA 4320

TTCCTGCCAA GCACCTTGAA TGGGAGGGGC CTCACAGAGG GCAGGGCCAG GGTCCAGCAG 4380

GGGTGGGGGG TTCCTGTTCT GCCCCTGCCC GTCCCCACCC AGTCTTGCCC TCCCATCCTC 4440

TCATCTATTC CCCCGCTGGA GACGGAAGAT CTTTTATTTT CTATTATTTA TAACTTCAGA 4500

CTTGGGCCCC CTGTTCTTTC TTTCCCATTA ACTTGAGTGA CCTGTGTGAG AGACAGACAG 4560

ATGCCCCACG AGGATGGCTG GACAAGGACT TTTACTTTTT ATTACATAAA AATATTAAAA 4620

AATAAATAAA AAAAATAAAA TTTTAAACTA ACTTAAAAAA AAAAAAAAAA AAAA 4674

( 2 ) INFORMATION FOR SEQ ID NO : 2 :

(i) SEQUENCE CHARACTERISTICS:

(A) LENGTH: 1051 amino acids

(B) TYPE: amino acid

(C) STRANDEDNESS: single

(D) TOPOLOGY: linear

(ii) MOLECULE TYPE: protein

(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2: Met Gly Trp Arg Pro Gly Arg Asp Ala Pro His Phe Pro Pro Ala Ala

1 5 10 15

Arg Pro Pro Pro Leu Pro Pro Arg Gly Ala Ser Ser Arg Arg Gly Phe 20 25 30 Leu Ser Pro Pro Pro Thr Pro Arg Gly Ser Pro Arg Pro Pro Thr Ala 35 40 45

Gly Pro Gly Cys Pro lie Pro Leu Ala Ser Arg Ala Leu Gin Arg Gly

50 55 60

Pro Ser Pro Gly Pro Leu Ser Pro Ser Leu Leu Ser Asn Ser Pro Ser 65 70 75 80

Gly Arg Cys His His Val Glu Ala Ser Ala Ala Thr Thr Ala Leu Pro

85 90 95

Ala Pro Ala Ala Ala Pro His Ala Thr Cys Arg Gly Pro Ser Ala Pro 100 105 110 Arg Gly His Ser Pro Pro Asn Gly Gly Leu Pro Gly Pro Leu Ala Thr 115 120 125

Ser Ala Ala Pro Pro Gly Pro Pro Ala Ala Ala Ser Pro Cys Leu Gly

130 135 140

Pro Val Ala Ala Ala Gly Ser Gly Leu Arg Arg Gly Ala Glu Gly lie 145 150 155 160

Leu Ala Pro Gin Pro Pro Pro Pro Gin Gin His Gin Glu Arg Pro Gly

165 170 175

Ala Ala Ala He Gly Ser Ala Arg Gly Gin Ser Thr Gly Lys Gly Pro 180 185 190 Pro Gin Ser Pro Val Phe Glu Gly Val Tyr Asn Asn Ser Arg Met Leu 195 200 205

His Phe Leu Thr Ala Val Val Gly Ser Thr Cys Asp Val Lys Val Lys

210 215 220

Asn Gly Thr Thr Tyr Glu Gly He Phe Lys Thr Leu Ser Ser Lys Phe 225 230 235 240

Glu Leu Ala Val Asp Ala Val His Arg Lys Ala Ser Glu Pro Ala Gly

245 250 255

Gly Pro Arg Arg Glu Asp He Val Asp Thr Met Val Phe Lys Pro Ser 260 265 270 Asp Val Met Leu Val His Phe Arg Asn Val Asp Phe Asn Tyr Ala Thr 275 280 285

Lys Asp Lys Phe Thr Asp Ser Ala He Ala Met Asn Ser Lys Val Asn

290 295 300

Gly Glu His Lys Glu Lys Val Leu Gin Arg Trp Glu Gly Gly Asp Ser 305 310 315 320

Asn Ser Asp Asp Tyr Asp Leu Glu Ser Asp Met Ser Asn Gly Trp Asp

325 330 335

Pro Asn Glu Met Phe Lys Phe Asn Glu Glu Asn Tyr Gly Val Lys Thr 340 345 350

Thr Tyr Asp Ser Ser Leu Ser Ser Tyr Thr Val Pro Leu Glu Lys Asp

355 360 365

Asn Ser Glu Glu Phe Arg Gin Arg Glu Leu Arg Ala Ala Gin Leu Ala 370 375 380

Arg Glu He Glu Ser Ser Pro Gin Tyr Arg Leu Arg He Ala Met Glu

385 390 395 400

Asn Asp Asp Gly Arg Thr Glu Glu Glu Lys His Ser Ala Val Gin Arg

405 410 415 Gin Gly Ser Gly Arg Glu Ser Pro Ser Leu Ala Ser Arg Glu Gly Lys

420 425 430

Tyr He Pro Leu Pro Gin Arg Val Arg Glu Gly Pro Arg Gly Gly Val

435 440 445

Arg Cys Ser Ser Ser Arg Gly Gly Arg Pro Gly Leu Ser Ser Leu Pro 450 455 460

Pro Arg Gly Pro His His Leu Asp Asn Ser Ser Pro Gly Pro Gly Ser

465 470 475 480

Glu Ala Arg Gly He Asn Gly Gly Pro Ser Arg Met Ser Pro Lys Ala

485 490 495 Gin Arg Pro Leu Arg Gly Ala Lys Thr Leu Ser Ser Pro Ser Asn Arg

500 505 510

Leu Leu Glu Asn Phe Cys Ser Thr Ser Ser Cys Ser Gly Pro Asp Val

515 520 525

Ser Pro Arg Ser Pro Lys Ser Ala Ala Pro Ala Pro He Ser Ala Ser 530 535 540

Cys Pro Glu Pro Pro He Gly Ser Ala Val Pro Thr Ser Ser Ala Ser

545 550 555 560

He Pro Val Thr Ser Ser Val Ser Asp Pro Gly Val Gly Ser He Ser

565 570 575 Pro Ala Ser Pro Lys He Ser Leu Ala Pro Thr Asp Val Lys Glu Leu

580 585 590

Ser Thr Lys Glu Pro Gly Arg Thr Leu Glu Pro Gin Glu Leu Ala Arg

595 600 605

He Ala Gly Lys Val Pro Gly Leu Gin Asn Glu Gin Lys Arg Phe Gin 610 615 620

Leu Glu Glu Leu Arg Lys Phe Gly Ala Gin Phe Lys Leu Gin Pro Ser 625 630 635 640

Ser Ser Pro Glu Asn Ser Leu Asp Pro Phe Pro Pro Arg He Leu Lys 645 650 655 Glu Glu Pro Lys Gly Lys Glu Lys Glu Val Asp Gly Leu Leu Thr Ser 660 665 670

Glu Pro Met Gly Ser Pro Val Ser Ser Lys Thr Glu Ser Val Ser Asp 675 680 685 Lys Glu Asp Lys Pro Pro Leu Ala Pro Ser Gly Gly Thr Glu Gly Pro

690 695 700

Glu Gin Pro Pro Pro Pro Cys Pro Ser Gin Thr Gly Ser Pro Pro Val 705 710 715 720 Gly Leu He Lys Gly Glu Asp Lys Asp Glu Gly Pro Val Ala Glu Gin

725 730 735

Val Lys Lys Ser Thr Leu Asn Pro Asn Ala Lys Glu Phe Asn Pro Thr

740 745 750

Lys Pro Leu Leu Ser Val Asn Lys Ser Thr Ser Thr Pro Thr Ser Pro 755 760 765

Gly Pro Arg Thr His Ser Thr Pro Ser He Pro Val Leu Thr Ala Gly

770 775 780

Gin Ser Gly Leu Tyr Ser Pro Gin Tyr He Ser Tyr He Pro Gin He 785 790 795 800 His Met Gly Pro Ala Val Gin Ala Pro Gin Met Tyr Pro Tyr Pro Val

805 810 815

Ser Asn Ser Val Pro Gly Gin Gin Gly Lys Tyr Arg Gly Ala Lys Gly

820 825 830

Ser Leu Pro Pro Gin Arg Ser Asp Gin His Gin Pro Ala Ser Ala Pro 835 840 845

Pro Met Met Gin Ala Ala Ala Ala Ala Gly Pro Pro Leu Val Ala Ala

850 855 860

Thr Pro Tyr Ser Ser Tyr He Pro Tyr Asn Pro Gin Gin Phe Pro Gly 865 870 875 880 Gin Pro Ala Met Met Gin Pro Met Ala His Tyr Pro Ser Gin Pro Val

885 890 895

Phe Ala Pro Met Leu Gin Ser Asn Pro Arg Met Leu Thr Ser Gly Ser

900 905 910

His Pro Gin Ala He Val Ser Ser Ser Thr Pro Gin Tyr Pro Ser Ala 915 920 925

Glu Gin Pro Thr Pro Gin Ala Leu Tyr Ala Thr Val His Gin Ser Tyr

930 935 940

Pro His His Ala Thr Gin Leu His Ala His Gin Pro Gin Pro Ala Thr 945 950 955 960 Thr Pro Thr Gly Ser Gin Pro Gin Ser Gin His Ala Ala Pro Ser Pro

965 970 975

Val Gin Val Pro Ala Met Gly Gly Ala Glu Trp Ser Trp Cys Arg Asn

980 985 990

Gly Trp Pro Glu Glu Gly He Glu Leu Gly Val He Ser Glu Trp Arg 995 1000 1005

Gly Leu Gly Ala Ser Glu Leu Leu Ala Cys Val Ala Leu Asn Leu Pro

1010 1015 1020

Leu Pro Ser Ser He Arg Arg Gly Arg Pro His Thr Trp Ala Val Asp 025 1030 1035 1040

Ser His Ser Arg He Cys Thr Thr Gin Gly Pro 1045 1050

Claims

What is claimed is:

1 An isolated polynucleotide compπsmg a nucleotide sequence that has at least 80% identity over its entire length to a nucleotide sequence encodmg the ataxιn-2 like gene polypeptide of SEQ ID NO 2. or a nucleotide sequence complementary to said isolated polynucleotide

2 The polynucleotide of claim 1 wherein said polynucleotide compπses the nucleotide sequence contamed SEQ ID NO 1 encodmg the ataxιn-2 like gene polypeptide of SEQ IDN02

3 The polynucleotide of claim 1 wherem said polynucleotide compπses a nucleotide sequence that is at least 80% identical to that of SEQ ID NO 1 over its entire length

4 The polynucleotide of claim 3 which is polynucleotide of SEQ ID NO 1

5 The polynucleotide of claim 1 which is DNA or RNA

6 A DNA or RNA molecule comprising an expression system, wherem said expression system is capable of producmg an ataxιn-2 like gene polypeptide comprising an ammo acid sequence, which has at least 80% identity with the polypeptide of SEQ ID NO 2 when said expression system is present m a compatible host cell

7 A host cell compπsmg the expression system of claim 6

8 A process for producmg an ataxιn-2 like gene polypeptide compnsmg cultuπng a host of claim 7 under conditions sufficient for the production of said polypeptide and recovermg the polypeptide from the culture

9 A process for producmg a cell which produces an ataxιn-2 like gene polypeptide thereof compπsmg transformmg or transfectmg a host cell with the expression system of claim 6 such that the host cell, under appropnate culture conditions, produces an ataxιn-2 like gene polypeptide 10 An ataxιn-2 like gene polypeptide compπsmg an ammo acid sequence which is at least 80% identical to the ammo acid sequence of SEQ ID NO 2 over its entire length

11 The polypeptide of claim 10 which comprises the ammo acid sequence of SEQ ID NO 2

12 An antibody immunospecific for the ataxιn-2 like gene polypeptide of claim 10

13 A method for the treatment of a subject m need of enhanced activity or expression of the ataxιn-2 like gene polypeptide of claim 10 compπsmg

(a) administering to the subject a therapeutically effective amount of an agomst to said polypeptide, and/or

(b) providing to the subject an isolated polynucleotide compπsmg a nucleotide sequence that has at least 80% identity to a nucleotide sequence encoding the ataxιn-2 like gene polypeptide of SEQ ID NO 2 over its entire length, or a nucleotide sequence complementary to said nucleotide sequence in a form so as to effect production of said polypeptide activity in vivo

14 A method for the treatment of a subject having need to inhibit activity or expression of the ataxιn-2 like gene polypeptide of claim 10 compπsmg (a) administering to the subject a therapeutically effective amount of an antagonist to said polypeptide. and/or

(b) administering to the subject a nucleic acid molecule that inhibits the expression of the nucleotide sequence encodmg said polypeptide. and/or

(c) administering to the subject a therapeutically effective amount of a polypeptide that competes with said polypeptide for its ligand. substrate , or receptor

15 A process for diagnosing a disease or a susceptibility to a disease m a subject related to expression or activity of the ataxιn-2 like gene polypeptide of claim 10 in a subject compπsmg (a) determining the presence or absence of a mutation m the nucleotide sequence encodmg said ataxιn-2 like gene polypeptide m the genome of said subject, and/or

(b) analyzing for the presence or amount of the ataxιn-2 like gene polypeptide expression m a sample deπved from said subject 16 A method for identifying compounds which inhibit (antagonize) or agonize the ataxιn-2 like gene polypeptide of claim 10 which compπses

(a) contacting a candidate compound with cells which express the ataxιn-2 like gene polypeptide (or cell membrane expressmg the ataxιn-2 like gene polypeptide) or respond to ataxιn-2 like gene polypeptide, and

(b) observing the binding, or stimulation or inhibition of a functional response, or comparing the abihty of the cells (or cell membrane) which were contacted with the candidate compounds with the same cells which were not contacted for ataxιn-2 like gene polypeptide activity

17 An agomst identified by the method of claim 16

18 An antagonist identified by the method of claim 16

19 A recombinant host cell produced by a method of Claim 9 or a membrane thereof expressmg an ataxιn-2 like gene polypeptide