HISTIDINE KINASE

Abstract: The invention provides Histidine Kinase polypeptides and polynucleotides encoding Histidine Kinase polypeptides and methods for producing such polypeptides by recombinant techniques. Also provided are methods for utilizing Histidine Kinase polypeptides to screen for antibacterial compounds.

Description: Histidine Kinase FIELD OF THE INVENTION This invention relates to newly identified polynucleotides and polypeptides. and their production and uses, as well as their variants, agonists and antagonists, and their uses In particular, the invention relates to novel polynucleotides and polypeptides of the Histidine Kinase family, hereinafter referred to as "Histidine Kinase" BACKGROUND OF THE INVENTION It is particularly preferred to employ Staphylococcal genes and gene products as targets for the development of antibiotics The Staphylococci make up a medically important genera of microbes They are known to produce two types of disease, invasive and toxigenic Invasive infections are characterized generally by abscess formation effecting both skin surfaces and deep tissues Staphylococcus aureus is the second leading cause of bacteremia in cancer patients Osteomyelitis, septic arthritis, septic thrombophlebitis and acute bactenal endocarditis are also relatively common There are at least three clinical conditions resulting from the toxigenic properties of Staphylococci The manifestation of these diseases result from the actions of exotoxins as opposed to tissue invasion and bacteremia These conditions include Staphylococcal food poisoning, scalded skin syndrome and toxic shock syndrome The frequency of Staphylococcus aureus infections has πsen dramatically in the past few decades This has been attributed to the emergence of multiply antibiotic resistant strains and an increasing population of people with weakened immune systems It is no longer uncommon to isolate Staphylococcus aureus strains which are resistant to some or all of the standard antibiotics This phenomenon has created a demand for both new aπti-microbial agents, vaccines, and diagnostic tests for this organism Many two component signal transduction systems (TCSTS) have been identified in bacteria (Stock, J B , Nrnfa, A J & Stock, A M (1989) Microbiol Rev 53, 450-490) These are involved m the bacterium's ability to monitor its surroundings and adapt to changes in its environment Several of these bacteπal TCSTS are mvolved m virulence and bacteπal pathogenesis within the host Histidine kmases are components of the TCSTS which autophosphorylate at a histidine residue The phosphate group is then transferred to the cognate response regulator The histidine kmases have five short conserved ammo acid sequences (Stock, J B . Ninfa, A J & Stock, A M (1989) Microbiol Rev 53, 450-490, Swanson, R V , Alex. L A & Simon, M I (1994) TIBS 19 485-491) These are the histidine residue, which is phosphorylated, followed after approximately 100 residues by a conserved asparagme residue After another 15 to 45 residues a DXGXG motif is found, followed by a FXXF motif after another 10-20 residues 10-20 residues further on another glycine motif, GXG is found The two glycine motifs are thought to be involved m nucleotide binding This family of histidine kmases mcludes KinA protein from Lactococcus lactis cremons (O'Connell-Motherway, M , Fitzgerald, G F & van Srnderen, D (1997) Appl Environ Microbiol 63 2454-2459) Response regulators are components of the TCSTS These protems are phosphorylated from histidine kmases and in turn once phosphorylated affect the response, often through a DNA bmdmg domam becoming activated The response regulators are characteπzed by a conserved N-terminal domain of approximately 100 amino acids The N- terminal domains of response regulators as well as retaining five functionally important residues, coπespondrng to the residues D12, D13, D57, T87, K109 m CheY (Matsumura, P , Rydel, J J , Linzmeier, R & Vacante, D (1984) J Bacteπol 160, 36-41), have conserved structural features (Volz, K (1993) Biochemistry 32, 11741-11753) The 3-dιmensιonal structures of CheY from Salmonella typhimurium (Stock, A M , Mottonen, J M , Stock, J B & Schutt, ,C E (1989) Nature, 337, 745-749) and Eschenchia coli (Volz, K & Matsumura, P (1991) J Biol Chem 266, 15511-15519) and the N-termmal domam of mtrogen regulatory protem C from S typhimurium (Volkman, B F , Nohaile, M J , Amy, N K , Kustu, S & Wemmer, D E (1995) Biochemistry, 34 1413-1424), are available, as well as the secondary structure of SpoOF from Bacillus subtilis (Feher, V A , Zapf, J W , Hoch, J A , Dahlquist, F W , Whiteley, J M & Cavanagh, J (1995) Protem Science, 4, 1801-1814) These structures have a (a/b)s fold Several structural residues are conserved between different response regulator sequences, specifically hydrophobic residues within the β-sheet hydrophobic core and sites from the a-hehces Among the processes regulated by TCSTS are production of virulence factors, motihty, antibiotic resistance and cell replication Inhibitors of TCSTS protems would prevent the bacteπum from establishmg and mamtainmg infection of the host by preventmg it from producmg the necessary factors for pathogenesis and thereby have utility in anti-bacteπal therapy Clearly, there exists a need for factors, such as the Histidme Kinase embodiments of the invention, that have a present benefit of bemg useful to screen compounds for antibiotic activity Such factors are also useful to determine their role in pathogenesis of infection, dysfunction and disease There is also a need for identification and characteπzation of such factors and their antagonists and agonists to find ways to prevent, ameliorate or correct such infection, dysfunction and disease Certain of the polypeptides of the invention possess ammo acid sequence homology to a known KinA from Lactococcus lactis cremons protem (O'Connell-Motherway, M , Fitzgerald, G F & van S deren, D (1997) Appl Environ Microbiol 63 2454-2459, U81166 from Genbank ) SUMMARY OF THE INVENTION It is an object of the invention to provide polypeptides that have been identified as novel Histidme Kinase polypeptides by homology between the ammo acid sequence set out in Table 1 [SEQ ID NO 2] and a known ammo acid sequence or sequences of other proteins such as KinA from Lactococcus lactis cremons protem It is a further object of the invention to provide polynucleotides that encode Histidme Kinase polypeptides, particularly polynucleotides that encode the polypeptide herein designated Histidme Kinase In a particularly preferred embodiment of the invention, the polynucleotide compπses a region encoding Histidme Kinase polypeptides compnsing a sequence set out in Table 1 [SEQ ID NO 1] which includes a full length gene, or a vanant thereof In another particularly prefened embodiment of the invention, there is a novel Histidine Kinase protem from Staphylococcus aureus compnsing the ammo acid sequence of Table 1 [SEQ ID NO 2], or a vanant thereof As a further aspect of the invention, there are provided isolated nucleic acid molecules encoding Histidme Kinase, particularly Staphylococcus aureus Histidme Kinase, including mRNAs, cDNAs, genomic DNAs Further embodiments of the invention mclude biologically, diagnostically, prophylactically, climcally or therapeutically useful vanants thereof, and compositions compnsing the same In accordance with another aspect of the invention, there is provided the use of a polynucleotide of the mvention for therapeutic or prophylactic purposes, m particular genetic immunization Among the particularly prefeπed embodiments of the mvention are naturally occurring allelic vanaπts of Histidme Kinase and polypeptides encoded thereby In another aspect of the mvention, there are provided novel polypeptides of Staphylococcus aureus refeπed to herein as Histidme Kinase as well as biologically, diagnostically, prophylactically, climcally or therapeutically useful vanants thereof, and compositions compπsmg the same Among the particularly prefeπed embodiments of the mvention are vanants of Histidme Kinase polypeptide encoded by naturally occurring alleles of the Histidme Kinase gene hi a prefeπed embodiment of the mvention, there are provided methods for producmg the aforementioned Histidine Kinase polypeptides In accordance with yet another aspect of the mvention, there are provided inhibitors to such polypeptides, useful as antibactenal agents, including, for example, antibodies In accordance with certain prefeπed embodiments of the mvention, there are provided products, compositions and methods for assessing Histidine Kinase expression, treating disease, assaying genetic vanation- and administering a Histidme Kinase polypeptide or polynucleotide to an organism to raise an immimological response against a bactena, especially a Staphylococcus aureus bactena In accordance with certain prefeπed embodiments of this and other aspects of the mvention, there are provided polynucleotides that hybndize to Histidme Kinase polynucleotide sequences, particularly under stringent conditions In certain prefeπed embodiments of the mvention, there are provided antibodies against Histidme Kinase polypeptides In other embodiments of the mvention, there are provided methods for identifying compounds which bmd to or otherwise interact with and inhibit or activate an activity of a polypeptide or polynucleotide of the mvention compnsing contacting a polypeptide or polynucleotide of the mvention with a compound to be screened under conditions to permit b dmg to or other interaction between the compound and the polypeptide or polynucleotide to assess the binding to or other interaction with the compound, such bmdmg or interaction being associated with a second component capable of providing a detectable signal m response to the bmdmg or interaction of the polypeptide or polynucleotide with the compound, and determining whether the compound binds to or otherwise interacts with and activates or inhibits an activity of the polypeptide or polynucleotide by detecting the presence or absence of a signal generated from the bmdmg or interaction of the compound with the polypeptide or polynucleotide In accordance with yet another aspect of the mvention, there are provided Histidme Kinase agonists and antagonists, preferably bactenostatic or bacteπocidal agonists and antagonists In a further aspect of the mvention, there are provided compositions compnsing a Histidme Kinase polynucleotide or a Histidme Kinase polypeptide for admmistration to a cell or to a multicellular organism Vanous changes and modifications within the spint and scope of the disclosed mvention will become readily apparent to those skilled m the art from reading the following descnptions and from reading the other parts of the present disclosure DESCRIPTION OF THE INVENTION The mvention relates to novel Histidme Kinase polypeptides and polynucleotides as descnbed in greater detail below In particular, the mvention relates to polypeptides and polynucleotides of a novel Histidme Kinase of Staphylococcus aureus, which is related by ammo acid sequence homology to KinA from Lactococcus lactis cremons polypeptide The mvention relates especially to Histidme Kinase havmg the nucleotide and ammo acid sequences set out m Table 1 as SEQ ID NO 1 and SEQ ID NO 2 respectively TABLE 1 Histidine Kinase Polynucleotide and Polypeptide Sequences (A) Sequences from Staphylococcus aureus Histidme Kinase polynucleotide sequence [SEQ ID NO 1] 5 ' -TAATTTAAAAAGCAACTATTGTATAGAAAAATACAAAATTTAAAATATATTACCTTATTA GAAAAAGTCGTAATATGAGGTGTACAAATGACGCAAATTTTAATAGTAGAAGATGAACAA AACTTAGCAAGATTTCTTGAATTGGAACTCACACATGAAAATTACAATGTGGACACAGAG TATGATGGACAAGACGGTTTAGATAAAGCGCTTAGCCATTACTATGATTTAATCATATTA GATTTAATGTTGCCGTCAATTAATGGCTTAGAAATTTGTCGCAAAATTAGACAACAACAA TCTACACCTATCATTATAATTACAGCGAAAAGTGATACGTATGACAAAGTTGCTGGGCTT GATTACGGTGCAGACGATTATATAGTTAAGCCGTTTGATATTGAAGAACTTTTAGCAAGA ATTCGTGCAATTTTACGTCGTCAGCCACAAAAGGATATTATCGATGTCAACGGTATTACA ATTGATAAGAACGCTTTTAAAGTGACGGTAAATGGCGCAGAAATTGAATTAACAAAAACA GAGTATGATTTACTATATCTTCTAGCTGAAAATAAAAACCATGTTATGCAACGGGAACAA ATTTTAAATCATGTATGGGGTTATAATAGTGAAGTAGAAACAAATGTCGTAGATGTTTAT ATAAGATATTTACGAAACAAGTTAAAACCATACGATCGTGACAAAATGATTGAAACAGTT CGTGGCGTTGGGTATGTGATACGATGACAAAACGTAAATTGCGCAATAACTGGATTATTG TTACCACGATGATTACGTTTGTCACGATATTTTTGTTTTGTTTAATTATTATTTTTTTCT TGAAAGATACACTGCATAATAGTGAGCTTGATGATGCAGAACGAAGCTCAAGCGATATTA ATAATTTATTTCATTCTAAGCCTGTTAAAGATATATCTGCATTAGACTTGAATGCATCTT TAGGTAATTTTCAAGAGATAATTATTTATGATGAGCATAATAATAAATTATTTGAGACAT CGAATGATAACACAGTGAGAGTTGAACCAGGTTATGAACACCGTTATTTTGACCGCGTAA TAAAAAAACGCTATAAAGGCATTGAATATTTAATTATTAAAGAACCAATTACAACGCAAG ATTTCAAAGGGTATAGCTTGTTAATTCATTCACTAGAAAATTATGATAACATCGTAAAAT CATTGTATATCATTGCGCTGGCATTTGGAGTGATTGCAACAATTATAACTGCCACAATCA GTTATGTATTTTCAACACAAATTACTAAACCGCTTGTCAGTTTATCAAATAAAATGATTG AGATTCGACGAGATGGTTTTCAAAATAAATTGCAATTAAATACAAATTATGAAGAAATAG ATAATTTAGCAAATACGTTTAATGAGATGATGAGCCAAATTGAAGAATCATTTAATCAAC AAAGACAATTTGTTGAAGATGCGTCACATGAATTACGAACACCATTACAAATTATTCAAG GTCATTTAAATTTGATTCAGCGATGGGGAAAAAAAGACCCAGCAGTATTAGAAGAATCGT TAAATATTTCTATTGAAGAAATGAATCGTATCATAAAATTAGTCGAAGAATTACTTGAAT TGACTAAAGGAGATGTAAATGACATTTCTTCTGAAGCGCAGACCGTGCATATTAATGATG AAATTCGCTCGCGAATACACTCATTAAAACAATTGCATCCTGATTATCAATTTGATACGG ATCTGACATCTAAAAATCTAGAAATTAAAATGAAACCTCATCAATTCGAACAATTATTTT TAATCTTTATTGATAATGCAATCAAATATGATGTGAAGAATAAGAAAATTAAAGTTAAGA CAAGGTTAAAAAATAAGCAAAAAATAATTGAAATTACAGATCATGGAATTGGTATTCCAG AGGAAGATCAAGATTTCATTTTTGATCGCTTTTATCGAGTGGATAAATCTCGTTCAAGAA GTCAAGGCGGTAATGGACTCGGATTATCTATTGCTCAAAAAATCATTCAATTAAACGGAG GATCGATTAAAATTAAAAGTGAAATTAATAAAGGAACAACGTTTAAAATCATATTTTAAT CATGTCTGAGACGTCAATCAAAGTCATAGGATCAATTTTTTAAGTACACATTAGCTGTGA CTAATGTATAAGAACAACTATAAAACAAATAAACAGTGGTT-3 ' (B) Staphylococcus aureus Histidine Kinase polypeptide sequence deduced from the polynucleotide sequence in this table [SEQ ID NO:2]. NH 2 -MTKRKLRNN IIV TMITFVTIFLFCLIIIFFLKDTLHNSELDDAERSSSDINNLFHSKP VKDISALDLNASLGNFQEIIIYDEHNNKLFETSNDNTVRVEPGYEHRYFDRVIKKRYKGI EYLIIKEPITTQDFKGYSLLIHSLENYDNIVKSLYIIALAFGVIATIITATISYVFSTQI TKPLVSLSNKMIEIRRDGFQNKLQLNTNYEEIDNLANTFNEMMSQIEESFNQQRQFVEDA SHELRTPLQIIQGHLNLIQR GKKDPAVLEESLNISIEEMNRIIKLVEELLELTKGDVND ISSEAQTVHINDEIRSRIHSLKQLHPDYQFDTDLTSKNLEIKMKPHQFEQLFLIFIDNAI KYDVKNKKIKVKTRLKNKQKIIEITDHGIGlPEEDQDFIFDRFYRVDKSRSRSQGGNGLG LSIAQKIIQLNGGSIKIKSEINKGTTFKIIF-COOH (C) Polynucleotide sequences comprising S.aureus response regulator cognate to the histidine kinase ofthe invention [SEQ ID NO 3] ATTTACGTTT TGTCATCGTA TCACATACCC AACGCCACGA ACTGTTTCAA TCATTTTGTC 60 ACGATCGTAT GGTTTTAACT TGTTTCGTAA ATATCTTATA TAAACATCTA CGACATTTGT 120 TTCTACTTCA CTATTATAAC CCCATACATG ATTTAAAATT TGTTCCCGTT GCATAACATG 180 GTTTTTATTT TCAGCTAGAA GATATAGTAA ATCATACTCT GTTTTTGTTA ATTCAATTTC 240 TGCGCCATTT ACCGTCACTT TAAAAGCGTT CTTATCAATT GTAATACCGT TGACATCGAT 300 AATATCCTTT TGTGGCTGAC GACGTAAAAT TGCACGAATT CTTGCTAAAA GTTCTTCAAT 360 ATCAAACGGC TTAACTATAT AATCGTCTGC ACCGTAATCA AGCCCAGCAA CTTTGTCATA 420 CGTATCACTT TTCGCTGTAA TTATAATGAT AGGTGTAGAT TGTTGTTGTC TAATTTTGCG 480 ACAAATTTCT AAGCCATTAA TTGACGGCAA CATTAAATCT AATATGATTA AATCATAGTA 540 ATGGCTAAGC GCTTTATCTA AACCGTCTTG TCCATCATAC TCTGTGTCCA CATTGTAATT 600 TTCATGTGTG AGTTCCAATT CAAGAAATCT TGCTAAGTTT TGTTCATCTT CTACTATTAA 660 AATTTGCGTC ATTTGTACAC CTCATATTAC GACTTTTTCT AATAAGGTAA TATATTTTAA 720 ATTTTGTATT TTTCTA 736 (D) Polypeptide sequence of Staphylococcus aureus response regulator cognate of the histidine kinase of the invention [SEQID NO 4] 1 MTQILIVEDE QNLARFLELE LTHENYNVDT EYDGQDGLDK ALSHYYDLII 51 LDLMLPSING LEICRKIRQQ QSTPIIIITA KSDTYDKVAG LDYGADDYIV 101 KPFDIEELLA RIRAILRRQP QKDIIDVNGI TIDKNAFKVT VNGAEIELTK 151 TEYDLLYLLA ENKNHVMQRE QILNHVWGYN SEVETNWDV YIRYLRNKLK 201 PYDRDKMIET VRGVGYVIR Deposited materials A deposit containing a Staphylococcus aureus WCUH 29 strain has been deposited with the National Collections of Industπal and Marine Bactena Ltd (herem "NCIMB"), 23 St Machar Dnve, Aberdeen AB2 1RN Scotland on 11 September 1995 and assigned NCIMB Deposit No 40771. and refeπed to as Staphylococcus aureus WCUH29 on deposit The Staphylococcus aureus strain deposit is refeπed to herem as "the deposited strain" or as "the DNA of the deposited strain " The deposited strain contains the full length Histidme Kinase gene The sequence of the polynucleotides contained m the deposited strain, as well as the ammo acid sequence of the polypeptide encoded thereby, are controlling in the event of any conflict with any descnption of sequences herem The deposit of the deposited strain has been made under the terms of the Budapest Treaty on the International Recognition of the Deposit of Micro-organisms for Purposes of Patent Procedure The strain will be irrevocably and without restπction or condition released to the public upon the issuance of a patent The deposited strain is provided merely as convemence to those of skill m the art and is not an admission that a deposit is required for enablement, such as that required under 35 U S C §112 A license may be required to make, use or sell the deposited strain, and compounds deπved therefrom, and no such license is hereby granted One aspect of the mvention there is provided an isolated nucleic acid molecule encoding a mature polypeptide expressible by the Staphylococcus aureus WCUH 29 strain contained in the deposited strain Further provided by the mvention are Histidme Kinase nucleotide sequences of the DNA in the deposited strain and ammo acid sequences encoded thereby Also provided by the mvention are Histidme Kinase polypeptide sequences isolated from the deposited strain and ammo acid sequences denved therefrom Polypeptides The polypeptides of the mvention include a polypeptide of Table 1 [SEQ ID NO 2] (in particular the mature polypeptide) as well as polypeptides and fragments, particularly those which have the biological activity of Histidme Kinase, and also those which have at least 70% identity to a polypeptide of Table 1 [SEQ ID NO l]or the relevant portion, preferably at least 80% identity to a polypeptide of Table 1 [SEQ ID NO 2and more preferably at least 90% siimlanty (more preferably at least 90% identity) to a polypeptide of Table 1 [SEQ ID NO 2] and still more preferably at least 95% similanty (still more preferably at least 95% identity) to a polypeptide of Table 1 [SEQ ED NO 2] and also include portions of such polypeptides with such portion of the polypeptide generally containing at least 30 ammo acids and more preferably at least 50 ammo acids The mvention also mcludes polypeptides of the formula X-(R 1 ) m -(R 2 )-(R 3 ) n -Y wherein, at the ammo terminus, X is hydrogen, and at the carboxyl terminus, Y is hydrogen or a metal, R\ and R3 are any ammo acid residue, m is an mteger between 1 and 1000 or zero, n is an mteger between 1 and 1000 or zero, and R 2 is an ammo acid sequence of the mvention, particularly an ammo acid sequence selected from Table 1 In the formula above R 2 is onented so that its amino terminal residue is at the left, bound to R and its carboxy terminal residue is at the nght, bound to R3 Any stretch of ammo acid residues denoted by either R group, where m and/or n is greater than 1, may be either a heteropolymer or a homopolymer, preferably a heteropolymer A fragment is a vanant polypeptide having an ammo acid sequence that entirely is the same as part but not all of the ammo acid sequence of the aforementioned polypeptides As with Histidme Kinase polypeptides fragments may be "free-standing," or compnsed within a larger polypeptide of which they form a part or region, most preferably as a single continuous region, a smgle larger polypeptide Prefeπed fragments clude, for example, truncation polypeptides havmg a portion of an ammo acid sequence of Table 1 [SEQ ED NO 2], or of vanants thereof, such as a continuous senes of residues that mcludes the ammo terminus, or a continuous senes of residues that mcludes the carboxyl terminus Degradation forms of the polypeptides of the mvention m a host cell, particularly a Staphylococcus aureus, are also prefeπed Further prefeπed are fragments characterized by structural or functional attributes such as fragments that compnse alpha-hehx and alpha-hehx forming regions, beta-sheet and beta-sheet-foiming regions, turn and turn-forrmng regions, coil and cod-forming regions, hydrophilic regions, hydrophobic regions, alpha amphipathic regions, beta amphipathic regions, flexible regions, surface-forming regions, substrate bmdmg region, and high antigemc index regions Also prefeπed are biologically active fragments which are those fragments that mediate activities of Histidme Kinase, including those with a similar activity or an improved activity, or with a decreased undesirable activity Also mcluded are those fragments that are antigemc or lmmunogenic in an animal, especially m a human Particularly prefeπed are fragments compnsing receptors or domains of enzymes that confer a function essential for viability of Staphylococcus aureus or the ability to initiate, or maintain cause disease m an individual, particularly a human Vanants that are fragments of the polypeptides of the mvention may be employed for producmg the coπesponding full-length polypeptide by peptide synthesis, therefore, these vanants may be employed as intermediates for producmg the full-length polypeptides of the mvention In addition to the standard smgle and tnple letter representations for ammo acids, the term "X" or "Xaa" may also be used m describing certain polypeptides of the mvention "X" and "Xaa" mean that any of the twenty naturally occunng amino acids may appear at such a designated position m the polypeptide sequence Polynucleotides Another aspect of the mvention relates to isolated polynucleotides, including the full length gene, that encode the Histidme Kinase polypeptide havmg a deduced ammo acid sequence of Table 1 [SEQ ID NO 2] and polynucleotides closely related thereto and vanants thereof Using the information provided herein, such as a polynucleotide sequence set out m Table 1 [SEQ ID NO 1], a polynucleotide of the mvention encoding Histidme Kinase polypeptide may be obtained us g standard cloning and screening methods, such as those for cloning and sequencmg chromosomal DNA fragments from bactena usmg Staphylococcus aureus WCUH 29 cells as starting matenal, followed by obtaining a full length clone For example, to obtain a polynucleotide sequence of the mvention, such as a sequence given m Table 1 [SEQ ID NO 1]. typically a library of clones of chromosomal DNA of Staphylococcus aureus WCUH 29 m E coli or some other suitable host is probed with a radiolabeled oligonucleotide, preferably a 17-mer or longer, denved from a partial sequence Clones carrying DNA identical to that of the probe can then be distinguished usmg stringent conditions By sequencmg the individual clones thus identified with sequencmg primers designed from the ongmal sequence it is then possible to extend the sequence m both directions to determme the full gene sequence Convemently, such sequencmg is performed usmg denatured double stranded DNA prepared from a plasmid clone Suitable techniques are descnbed by Maniatis, T , Fntsch, E F and Sambrook et al , MOLECULAR CLONING, A LABORATORY MANUAL, 2nd Ed , Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York (1989) (see m particular Screening By Hybndization 1 90 and Sequencmg Denatured Double-Stranded DNA Templates 13 70) Illustrative of the mvention, the polynucleotide set out m Table 1 [SEQ ID NO 1] was discovered m a DNA library denved from Staphylococcus aureus WCUH 29 The DNA sequence set out in Table 1 [SEQ ID NO 1] contains an open reading frame encoding a protem having about the number of ammo acid residues set forth m Table 1 [SEQ ID NO 2] with a deduced molecular weight that can be calculated usmg amino acid residue molecular weight values well known in the art The polynucleotide of SEQ ID NO 1, between nucleotide number 744 and the stop codon which begins at nucleotide number 2097 of SEQ ID NO 1, encodes the polypeptide of SEQ ID NO 2 Histidme Kinase of the mvention is structurally related to other proteins of the Histidme Kinase family The mvention provides a polynucleotide sequence identical over its entire length to a coding sequence m Table 1 [SEQ ID NO 1] Also provided by the mvention is the coding sequence for the mature polypeptide or a fragment thereof, by itself as well as the coding sequence for the mature polypeptide or a fragment in reading frame with other coding sequence, such as those encoding a leader or secretory sequence, a pre-, or pro- or prepro- protem sequence The polynucleotide may also contain non-coding sequences, mcludmg for example, but not limited to non-coding 5' and 3' sequences, such as the transcnbed, non-translated sequences, termination signals, nbosome binding sites, sequences that stabilize mRNA, nitrons, polyadenylation signals, and additional coding sequence which encode additional ammo acids For example, a marker sequence that facilitates purification of the fused polypeptide can be encoded In certain embodiments of the mvention, the marker sequence is a hexa-histidme peptide, as provided m the pQE vector (Qiagen, Inc ) and descnbed m Gentz et al , Proc Natl Acad Sci , USA 86 821-824 (1989), or an HA tag (Wilson et al , Cell 37 161 (1984) Polynucleotides of the mvention also clude, but are not limited to, polynucleotides compnsing a structural gene and its naturally associated sequences that control gene expression A prefeπed embodiment of the mvention is a polynucleoti.de of compnsing nucleotide 744 to the nucleotide immediately upstream of or mcludmg nucleotide 2097 set forth in SEQ ID NO 1 of Table 1, both of which encode the Histidme Kinase polypeptide The mvention also mcludes polynucleotides of the formula X-(R 1 ) m -(R 2 )-(R 3 ) n -Y wherein, at the 5' end of the molecule, X is hydrogen or together with Y defines a covalent bond, and at the 3' end of the molecule, Y is hydrogen or a metal or together with X defines the covalent bond, each occurance of Rj and R3 is mdependently any nucleic acid residue, m is an mteger between 1 and 3000 or zero , n is an mteger between 1 and 3000 or zero, and R 2 is a nucleic acid sequence of the invention, particularly a nucleic acid sequence selected from Table 1 In the polynucleotide formula above R 2 is oriented so that its 5' end residue is at the left, bound to R and its 3' end residue is at the nght, bound to R3 Any stretch of nucleic acid residues denoted by either R group, where m and/or n is greater than 1. may be either a heteropolymer or a homopolymer, preferably a heteropolymer Where m a prefeπed embodiment. X and Y together define a covalent bond, the polynucleotide of the above formula is a closed, circular polynucleotide, which can be a double-stranded polynucleotide wherein the formula shows a first strand to which the second strand is complementary In another prefeπed embodiment m and/or n is an mteger between 1 and 1000. It is most prefeπed that the polynucleotides of the inventions are denved from Staphylococcus aureus, however, they may preferably be obtained from organisms of the same taxonomic genus They may also be obtained, for example, from orgamsims of the same taxonomic family or order The term "polynucleotide encoding a polypeptide" as used herem encompasses polynucleotides that mclude a sequence encoding a polypeptide of the mvention, particularly a bactenal polypeptide and more particularly a polypeptide of the Staphylococcus aureus Histidme Kinase havmg an ammo acid sequence set out m Table 1 [SEQ ID NO 2] The term also encompasses polynucleotides that mclude a smgle continuous region or discontinuous regions encoding the polypeptide (for example, interrupted by integrated phage or an insertion sequence or editing) together with additional regions, that also may contain coding and/or non-codmg sequences The mvention further relates to vanants of the polynucleotides descnbed herem that encode for vanants of the polypeptide havmg a deduced ammo acid sequence of Table 1 [SEQ ID NO 2] Vanants that are fragments of the polynucleotides of the mvention may be used to synthesize full- length polynucleotides of the mvention Further particularly prefeπed embodiments are polynucleotides encoding Histidme Kinase vanants, that have the ammo acid sequence of Histidme Kinase polypeptide of Table 1 [SEQ ED NO 2] in which several, a few, 5 to 10, 1 to 5, 1 to 3, 2, 1 or no ammo acid residues are substituted, deleted or added, m any combination Especially prefeπed among these are silent substitutions, additions and deletions, that do not alter the properties and activities of Histidme Kinase Further prefeπed embodiments of the mvention are polynucleotides that are at least 70% identical over their entire length to a polynucleotide encoding Histidme Kinase polypeptide havmg an ammo acid sequence set out m Table 1 [SEQ ID NO 2], and polynucleotides that are complementary to such polynucleotides Alternatively, most highly prefeπed are polynucleotides that compπse a region that is at least 80% identical over its entire length to a polynucleotide encoding Histidme Kinase polypeptide and polynucleotides complementary thereto hi this regard, polynucleotides at least 90%o identical over their entire length to the same are particularly prefeπed, and among these particularly prefeπed polynucleotides, those with at least 95% are especially prefeπed Furthermore, those with at least 97% are highly prefeπed among those with at least 95%, and among these those with at least 98% and at least 99% are particularly highly prefeπed, with at least 99% bemg the more prefeπed Prefeπed embodiments are polynucleotides that encode polypeptides that retain substantially the same biological function or activity as the mature polypeptide encoded by a DNA ofTable l [SEQ ID N0 1] The mvention further relates to polynucleotides that hybndize to the herem above-descnbed sequences In this regard, the mvention especially relates to polynucleotides that hybndize under stringent conditions to the herem above-descnbed polynucleotides As herem used, the terms "stringent conditions" and "stringent hybndization conditions" mean hybndization will occur only if there is at least 95% and preferably at least 97% identity between the sequences An example of stringent hybndization conditions is overnight incubation at 42°C m a solution compnsmg 50% formamide, 5x SSC (150mM NaCl, 15mM tnsodmm citrate), 50 mM sodium phosphate (pH7 6), 5x Denhardt's solution, 10% dextran sulfate, and 20 micrograms/ml denatured, sheared salmon sperm DNA, followed by washing the hybndization support m 0 lx SSC at about 65 °C Hybndization and wash conditions are well known and exemplified m Sambrook, et al , Molecular Cloning A Laboratory Manual, Second Edition, Cold Spring Harbor, N Y , (1989), particularly Chapter 11 therein The mvention also provides a polynucleotide consistmg essentially of a polynucleotide sequence obtamable by screenmg an appropnate library contammg the complete gene for a polynucleotide sequence set forth m SEQ ID NO 1 under stringent hybndization conditions with a probe havmg the sequence of said polynucleotide sequence set forth m SEQ ID NO 1 or a fragment thereof, and isolating said DNA sequence Fragments useful for obtaining such a polynucleotide mclude, for example, probes and primers descnbed elsewhere herem As discussed additionally herem regarding polynucleotide assays of the mvention, for instance, polynucleotides of the mvention as discussed above, may be used as a hybndization probe for RNA, cDNA and genomic DNA to isolate full-length cDNAs and genomic clones encoding Histidme Kinase and to isolate cDNA and genomic clones of other genes that have a high sequence similanty to the Histidme Kinase gene Such probes generally will compπse at least 15 bases Preferably, such probes will have at least 30 bases and may have at least 50 bases Particularly prefeπed probes will have at least 30 bases and will have 50 bases or less For example, the coding region of the Histidme Kinase gene may be isolated by screening usmg a DNA sequence provided m Table 1 [SEQ ID NO 1] to synthesize an oligonucleotide probe A labeled oligonucleotide havmg a sequence complementary to that of a gene of the mvention is then used to screen a library of cDNA, genomic DNA or mRNA to determine which members of the library the probe hybndizes to The polynucleotides and polypeptides of the mvention may be employed, for example, as research reagents and matenals for discovery of treatments of and diagnostics for disease, particularly human disease, as further discussed herem relatmg to polynucleotide assays Polynucleotides of the mvention that are oligonucleotides denved from the sequences of Table 1 [SEQ ID NOS 1 or 2] may be used m the processes herem as described, but preferably for PCR, to determme whether or not the polynucleotides identified herem m whole or m part are transcribed m bactena m infected tissue It is recognized that such sequences will also have utility in diagnosis of the stage of infection and type of infection the pathogen has attained The mvention also provides polynucleotides that may encode a polypeptide that is the mature protem plus additional ammo or carboxyl-terminal ammo acids, or ammo acids intenor to the mature polypeptide (when the mature form has more than one polypeptide chain, for instance) Such sequences may play a role in processing of a protem from precursor to a mature form, may allow protem transport, may lengthen or shorten protem half-life or may facilitate manipulation of a protem for assay or production, among other things As generally is the case in vivo, the additional ammo acids may be processed away from the mature protem by cellular enzymes A precursor protein, having the mature form of the polypeptide fused to one or more prosequences may be an inactive form of the polypeptide When prosequences are removed such inactive precursors generally are activated Some or all of the prosequences may be removed before activation Generally, such precursors are called proproteins In addition to the standard A, G, C, T/U representations for nucleic acid bases, the term "N" may also be used m descnbmg certam polynucleotides of the mvention "N" means that any of the four DNA or RNA bases may appear at such a designated position m the DNA or RNA sequence, except it is prefeπed that N is not a base that when taken m combmation with adjacent nucleotide positions, when read m the coπect readmg frame would have the effect of generatmg a premature termination codon m such readmg frame In sum, a polynucleotide of the mvention may encode a mature protein, a mature protem plus a leader sequence (which may be refeπed to as a preprotein), a precursor of a mature protem havmg one or more prosequences that are not the leader sequences of a preprotein, or a preproprotein- which is a precursor to a proprotein, havmg a leader sequence and one or more prosequences, which generally are removed dunng processmg steps that produce active and mature forms of the polypeptide Vectors, host cells, expression The mvention also relates to vectors that compnse a polynucleotide or polynucleotides of the mvention, host cells that are genetically engineered with vectors of the mvention and the production of polypeptides of the mvention by recombinant techniques Cell-free translation svstems can also be employed to produce such proteins usmg RNAs denved from the DNA constructs of the mvention For recombinant production, host cells can be genetically engineered to incorporate expression systems or portions thereof or polynucleotides of the mvention Introduction of a polynucleotide mto the host cell can be effected by methods descnbed m 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, DEAE-dextran mediated transfection, transvection, microinjection, catiomc hpid- mediated transfection, electroporation, transduction, scrape loading, ballistic introduction and infection Representative examples of appropnate hosts mclude bactenal cells, such as streptococci, staphylococci, eπterococci E coli, streptomyces and Bacillus subtilis 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, 293 and Bowes melanoma cells, and plant cells A great vanety of expression systems can be used to produce the polypeptides of the mvention Such vectors mclude, among others, chromosomal, episomal and virus-denved vectors, e g , vectors denved from bacteπal plasrmds, 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, fowl pox viruses, pseudorabies viruses and retroviruses, and vectors denved from combinations thereof, such as those denved from plasmid and bactenophage genetic elements, such as cosmids and phagemids The expression system constructs may contain control regions that regulate as well as engender expression Generally, any system or vector suitable to maintain, propagate or express polynucleotides and/or to express a polypeptide m a host may be used for expression m this regard The appropnate DNA sequence may be inserted mto the expression system by any of a vanety of well-known and routine techniques, such as, for example, those set forth m Sambrook et al , MOLECULAR CLONING, A LABORATORY MANUAL, (supra) For secretion of the translated protem mto the lumen of the endoplasmic reticulum, mto the penplasmic space or mto the extracellular environment, appropπate secretion signals may be incorporated mto the expressed polypeptide These signals may be endogenous to the polypeptide or they may be heterologous signals Polypeptides of the mvention can be recovered and purified from recombinant cell cultures by well-known methods mcludmg ammonium sulfate 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 purification Well known techniques for refolding protem may be employed to regenerate active conformation when the polypeptide is denatured dunng isolation and or purification Diagnostic Assays This mvention is also related to the use of the Histidme Kinase polynucleotides of the mvention for use as diagnostic reagents Detection of Histidme Kinase m a eukaryote, particularly a mammal, and especially a human, will provide a diagnostic method for diagnosis of a disease Eukaryotes (herein also "mdιvιdual(s)"), particularly mammals, and especially humans, particularly those infected or suspected to be infected with an organism compnsmg the Histidme Kinase gene may be detected at the nucleic acid level by a vanety of techniques Nucleic acids for diagnosis may be obtained from an infected individual's cells and tissues, such as bone, blood, muscle, cartilage, and skin Genomic DNA may be used directly for detection or may be amplified enzymatically by usmg PCR or other amplification technique pnor to analysis RNA, cDNA and genomic DNA may also be used m the same ways Using amplification, characterization of the species and strain of prokaryote present m an individual, may be made by an analysis of the genotype of the prokaryote gene Deletions and insertions can be detected by a change m size of the amplified product m companson to the genotype of a reference sequence Point mutations can be identified by hybndizmg amplified DNA to labeled Histidine Kinase polynucleotide 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 m the electrophoretic mobihty of the DNA fragments m gels, with or without denaturing agents, or by direct DNA sequencmg See, e g , Myers et al , Science 230 1242 (1985) Sequence changes at specific locations also may be revealed by nuclease protection assays, such as RNase and SI protection or a chemical cleavage method See, e g , Cotton et al . Proc Natl Acad Sci , USA, 85 4397-4401 (1985) Cells carrying mutations or polymorphisms m the gene of the mvention may also be detected at the DNA level by a vanety of techniques, to allow for serotyping, for example For example, RT- PCR can be used to detect mutations It is particularly prefeπed to used RT-PCR m conjunction with automated detection systems, such as, for example, GeneScan RNA, cDNA or genomic DNA may also be used for the same purpose, PCR or RT-PCR As an example, PCR primers complementary to a nucleic acid encodmg Histidme Kinase can be used to identify and analyze mutations Examples of representative primers are shown below m Table 2 Table 2 Primers for amplification of Histidine Kinase polynucleotides SEQ ID NO PRIMER SEQUENCE 5 5 ' -ATGACAAAACGTAAATTGCGCAATAAC-3 ' 6 5 ' -AAATATGATTTTAAACGTTGTTCC-3 ' The mvention also mcludes primers of the formula X-(R 1 ) m -(R 2 )-(R 3 ) n N wherein, at the 5' end of the molecule, X is hydrogen, and at the 3' end of the molecule, Y is hydrogen or a metal, R\ and R3 is any nucleic acid residue, m is an mteger between 1 and 20 or zero , n is an mteger between 1 and 20 or zero, and R is a primer sequence of the mvention, particularly a primer sequence selected from Table 2 In the polynucleotide formula above R is onented so that its 5' end residue is at the left, bound to R\ and its 3' end residue is at the nght, bound to R3 Any stretch of nucleic acid residues denoted by either R group, where m and/or n is greater than 1, may be either a heteropolymer or a homopolymer, preferably a heteropolymer bemg complementary to a region of a polynucleotide of Table 1 In a prefeπed embodiment m and or n is an mteger between 1 and 10. The mvention further provides these primers with 1, 2, 3 or 4 nucleotides removed from the 5' and/or the 3' end These primers may be used for. among other things, amplifying Histidme Kinase DNA isolated from a sample denved from an individual The primers may be used to amplify the gene isolated from an infected individual such that the gene may then be subject to vanous techniques for elucidation of the DNA sequence In this way, mutations m the DNA sequence may be detected and used to diagnose infection and to serotype and/or classify the infectious agent The mvention further provides a process for diagnosing, disease, preferably bacterial infections, more preferably infections by Staphylococcus aureus, compnsmg determining from a sample denved from an individual a mcreased level of expression of polynucleotide havmg a sequence of Table 1 [SEQ ID NO 1] Increased or decreased expression of Histidme Kinase polynucleotide can be measured usmg any on of the methods well known m the art for the quantation of polynucleotides, such as, for example, amplification, PCR, RT-PCR, RNase protection, Northern blotting and other hybndization methods In addition, a diagnostic assay m accordance with the mvention for detecting over- expression of Histidme Kinase protem compared to normal control tissue samples may be used to detect the presence of an infection, for example Assay techniques that can be used to determme levels of a Histidme Kinase protein, m 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 Antibodies The polypeptides of the mvention or vanants thereof, or cells expressmg them can be used as an immunogen to produce antibodies immunospecific for such polypeptides "Antibodies" as used herem mcludes monoclonal and polyclonal antibodies, chimeπc, single chain, simiamzed antibodies and humanized antibodies, as well as Fab fragments, mcludmg the products of an Fab immunoglobuhn expression library Antibodies generated against the polypeptides of the mvention can be obtained by administering the polypeptides or epitope-beanng fragments, analogues or cells to an animal, preferably a nonhuman, usmg routine protocols For preparation of monoclonal antibodies, any technique known m the art that provides antibodies produced by continuous cell hne cultures can be used Examples mclude vanous techniques, such as those m Kohler, G and M-lste-n, C , Nature 256 495-497 (1975), Kozbor et al , Immunology Today 4 72 (1983), Cole et al , pg 77-96 in MONOCLONAL ANTIBODIES AND CANCER THERAPY, Alan R Liss, Inc (1985) Techniques for the production of smgle chain antibodies (U S Patent No 4,946,778) can be adapted to produce smgle chain antibodies to polypeptides of this mvention Also, transgenic mice, or other organisms such as other mammals, may be used to express humanized antibodies Alternatively phage display technology may be utilized to select antibody genes with bmdmg activities towards the polypeptide either from repertoires of PCR amplified v-genes of lymphocytes from humans screened for possessmg anti-Histidine Kinase or from naive hbranes (McCafferty, J et al , (1990), Nature 348, 552-554, Marks, J et al , (1992) Biotechnology 10. 779-783) The affinity of these antibodies can also be improved by cham shuffling (Clackson, T et al , (1991) Nature 352, 624-628) If two antigen bmdmg domains are present each domain may be directed agamst a different epitope - termed 'bispecific' antibodies The above-descnbed antibodies may be employed to isolate or to identify clones expressmg the polypeptides to purify the polypeptides by affinity chromatography Thus, among others, antibodies against Histidme Kinase-polypeptide may be employed to treat infections, particularly bactenal infections Polypeptide vanants mclude antigemcally, epitopically or lmmunologically equivalent variants that form a particular aspect of this mvention The term "antigemcally equivalent denvative" as used herem encompasses a polypeptide or its equivalent which will be specifically recognized by certam antibodies which, when raised to the protem or polypeptide accordmg to the invention, interfere with the immediate physical mteraction between pathogen and mammalian host The term "lmmunologically equivalent denvative" as used herem encompasses a peptide or its eqmvalent which when used m a smtable formulation to raise antibodies m a vertebrate, the antibodies act to interfere with the immediate physical mteraction between pathogen and mammalian host The polypeptide, such as an antigemcally or lmmunologically equivalent denvative or a fusion protem thereof is used as an antigen to immunize a mouse or other animal such as a rat or chicken The fusion protem may provide stability to the polypeptide The antigen may be associated, for example by conjugation, with an lmmunogenic earner protem for example bovme serum albumin (BSA) or keyhole limpet haemocyanin (KLH) Alternatively a multiple antigenic peptide compnsmg multiple copies of the protem or polypeptide, or an antigemcally or lmmunologically equivalent polypeptide thereof may be sufficiently antigemc to improve immunogenicity so as to obviate the use of a earner Preferably, the antibody or vanant thereof is modified to make it less lmmunogemc m the mdividual For example, if the mdividual is human the antibody may most preferably be "humanized", where the complimentaπty determining regιon(s) of the hybndoma-denved antibody has been transplanted mto a human monoclonal antibody , for example as descnbed m Jones, P et al (1986), Nature 321, 522-525 or Tempest et al , (1991) Biotechnology 9, 266-273 The use of a polynucleotide of the mvention m genetic immunization will preferably employ a suitable delivery method such as direct injection of plasmid DNA mto muscles (Wolff et al , Hum Mol Genet 1992, 1 363, Manthorpe et al , Hum Gene Ther 1963 4, 419), delivery of DNA complexed with specific protem earners (Wu et al , J Biol Chem 1989 264,16985), coprecipitation of DNA with calcium phosphate (Benvemsty & Reshef, PNAS USA, 1986 83,9551), encapsulation of DNA m various forms of liposomes (Kaneda et al , Science 1989 243.375), particle bombardment (Tang et al , Nature 1992, 356 152, Eisenbraun et al , DNA Cell Biol 1993, 12 791) and in vivo infection usmg cloned retroviral vectors (Seeger et al , PNAS USA 1984 81,5849) Antagonists and agonists - assays and molecules Polypeptides of the mvention may also be used to assess the bmdmg of small molecule substrates and ligands in, for example, cells, cell-free preparations, chemical hbranes, and natural product mixtures These substrates and ligands may be natural substrates and ligands or may be structural or functional mimetics See, e g , Cohgan et al , Current Protocols in Immunology 1(2) Chapter 5 (1991) The mvention also provides a method of screenmg compounds to identify those which enhance (agonist) or block (antagonist) the action of Histidme Kinase polypeptides or polynucleotides, particularly those compounds that are bactenostatic and/or bactenocidal The method of screening may mvolve high-throughput techniques For example, to screen for agonists or antagoists, a synthetic reaction mix, a cellular compartment, such as a membrane, cell envelope or cell wall, or a preparation of any thereof, compnsmg Histidme Kinase polypeptide and a labeled substrate or hgand of such polypeptide is incubated m the absence or the presence of a candidate molecule that may be a Histidme Kinase agonist or antagonist The ability of the candidate molecule to agonize or antagonize the Histidme Kinase polypeptide is reflected m decreased bmdmg of the labeled ligand or decreased production of product from such substrate Molecules that bmd gratuitously, . e , without mducmg the effects of Histidme Kinase polypeptide are most likely to be good antagonists Molecules that bmd well and mcrease the rate of product production from substrate are agonists Detection of the rate or level of production of product from substrate may be enhanced by using a reporter system Reporter systems that may be useful m this regard mclude but are not limited to colonmetnc labeled substrate converted mto product, a reporter gene that is responsive to changes m Histidme Kinase polynucleotide or polypeptide activity, and bmdmg assays known m the art Another example of an assay for Histidme Kinase antagonists is a competitive assay that combmes Histidme Kinase and a potential antagonist with Histidme K-nase-binding molecules, recombinant Histidme Kinase bmdmg molecules, natural substrates or hgands. or substrate or ligand mimetics, under appropnate conditions for a competitive inhibition assay Histidine Kinase can be labeled, such as by radioactivity or a colonmetnc compound, such that the number of Histidme Kinase molecules bound to a bmdmg molecule or converted to product can be determmed accurately to assess the effectiveness of the potential antagonist Potential antagonists mclude small organic molecules, peptides, polypeptides and antibodies that bmd to a polynucleotide or polypeptide of the mvention and thereby inhibit or extinguish its activity Potential antagonists also may be small organic molecules, a peptide, a polypeptide such as a closely related protem or antibody that bmds the same sites on a bmdmg molecule, such as a bmdmg molecule, without mducmg Histidme Kinase-induced activities, thereby preventing the action of Histidine Kinase by excluding Histidme Kinase from bmdmg Potential antagonists mclude a small molecule that bmds to and occupies the bmdmg site of the polypeptide thereby preventing bmdmg to cellular binding molecules, such that normal biological activity is prevented Examples of small molecules mclude but are not limited to small organic molecules, peptides or peptide-hke molecules Other potential antagonists mclude antisense molecules (see Okano, J Neurochem 56 560 (1991), OUGODEOXYNUCLEOTIDES AS ANTISENSE INHIBITORS OF GENE EXPRESSION, CRC Press, Boca Raton, FL (1988), for a descnption of these molecules) Prefeπed potential antagonists mclude compounds related to and vanants of Histidme Kinase Each of the DNA sequences provided herem may be used m the discovery and development of antibactenal compounds The encoded protem, upon expression, can be used as a target for the screenmg of antibactenal drugs Additionally, the DNA sequences encodmg the ammo terminal regions of the encoded protem or Shme-Delgarno or other translation facilitating sequences of the respective mRNA can be used to construct antisense sequences to control the expression of the coding sequence of mterest The mvention also provides the use of the polypeptide, polynucleotide or inhibitor of the mvention to interfere with the initial physical mteraction between a pathogen and mammalian host responsible for sequelae of infection In particular the molecules of the invention may be used in the prevention of adhesion of bactena, m particular gram positive bactena, to mammalian extracellular matnx protems on in-dwelling devices or to extracellular matrix protems m wounds, to block Histidme Kinase protem-mediated mammalian cell mvasion by, for example, initiating phosphorylation of mammalian tyrosme kmases (Rosenshine et al , Infect Immun 60 2211 (1992), to block bactenal adhesion between mammalian extracellular matnx protems and bactenal Histidme Kinase protems that mediate tissue damage and, to block the normal progression of pathogenesis m infections initiated other than by the implantation of in-dwelling devices or by other surgical techniques This mvention provides a method of screenmg drugs to identify those which interfere with I) the mteraction of the histidme kinase with a response regulator, the method compnsmg mcubatmg the histidme kmase with response regulator in the presence of the drug and measuring the ability of the drug to block this mteraction, and/or n) the ability of the histidme kinase to autophosphorylate, the method compnsmg mcubatmg the histidme kmase with the drug and measuring the ability of the drug to prevent autophosphorylation The response regulator is preferably the cognate response regulator of the histidme kmase, or another response regulator which is capable of usmg the histidme kmase as a substrate, and is preferably from Staphylococcus aureus or another microorganism e g Bacillus Polypeptide and polynucleotide sequences of the cognate response regulator of the Histidme kmase of the mvention are set forth m Table 1 (C and D) This novel response regulator shows 42% identity to the ResD response regulator protem from Bacillus subtilis The antagonists and agonists of the mvention may be employed, for instance, to inhibit and treat diseases Hehcobacter pylori (herem H pylori) bactena infect the stomachs of over one-third of the world's population causmg stomach cancer, ulcers, and gastntis (International Agency for Research on Cancer (1994) Schistosomes, Liver Flukes and Hehcobacter Pylon (International Agency for Research on Cancer, Lyon, France, http //www uicc ch/ecp/ecp2904 htm) Moreover, the international Agency for Research on Cancer recently recognized a cause-and- effect relationship between H pylori and gastric adenocarcmoma, classifying the bacterium as a Group I (definite) carcmogen Prefened antimicrobial compounds of the mvention (agomsts and antagonists of Histidme Kinase) found usmg screens provided by the mvention, particularly broad-spectrum antibiotics, should be useful m the treatment of H pylon mfection Such treatment should decrease the advent of H /rμ/σrz -induced cancers, such as gastrointestinal carcmoma Such treatment should also cure gastric ulcers and gastntis Vaccines Another aspect of the mvention relates to a method for mducmg an immunological response m an mdividual, particularly a mammal which compnses inoculating the mdividual with Histidme Kinase, or a fragment or vanant thereof, adequate to produce antibody and/ or T cell immune response to protect said mdividual from infection, particularly bactenal infection and most particularly Staphylococcus aureus infection Also provided are methods whereby such immunological response slows bacterial replication Yet another aspect of the mvention relates to a method of mducmg immunological response m an mdividual which compnses delivering to such mdividual a nucleic acid vector to direct expression of Histidine Kinase, or a fragment or a vanant thereof, for expressmg Histidine Kinase, or a fragment or a vanant thereof in vivo m order to induce an immunological response, such as, to produce antibody and or T cell immune response, mcludmg, for example, cytokme-producmg T cells or cytotoxic T cells, to protect said mdividual from disease, whether that disease is already established within the mdividual or not One way of administering the gene is by acceleratmg it mto the desired cells as a coatmg on particles or otherwise Such nucleic acid vector may compnse DNA, RNA, a modified nucleic acid, or a DNA/RNA hybnd A further aspect of the mvention relates to an immunological composition which, when mtroduced mto an mdividual capable or havmg mduced within it an immunological response, mduces an immunological response m such mdividual to a Histidme Kinase or protem coded therefrom, wherem the composition compnses a recombinant Histidme Kinase or protem coded therefrom compnsmg DNA which codes for and expresses an antigen of said Histidme Kinase or protem coded therefrom The unmunological response may be used therapeutically or prophylactically and may take the form of antibody immunity or cellular immunity such as that aπsmg from CTL or CD4+ T cells A Histidme Kinase polypeptide or a fragment thereof may be fused with co-protem which may not by itself produce antibodies, but is capable of stabilizing the first protem and producmg a fused protem which will have immunogenic and protective properties Thus fused recombinant protem, preferably further comprises an antigemc co-protem, such as lrpoprotein D from Hemophilus influenzae, Glutathione-S-transferase (GST) or beta-galactosidase, relatively large co-protems which solubihze the protem and facilitate production and punfication thereof Moreover, the co-protem may act as an adjuvant m the sense of providmg a generalized stimulation of the immune system The co-protein may be attached to either the am o or carboxy terminus of the first protem Provided by this invention are compositions, particularly vaccine compositions, and methods compnsing the polypeptides or polynucleotides of the mvention and lmmunostimulatory DNA sequences, such as those descnbed m Sato, Y et al Science 273 352 (1996) Also, provided by this mvention are methods usmg the descnbed polynucleotide or particular fragments thereof which have been shown to encode non-vanable regions of bactenal cell surface protems m DNA constructs used in such genetic immunization experiments m animal models of infection with Staphylococcus aureus will be particularly useful for identifying protem epitopes able to provoke a prophylactic or therapeutic immune response It is believed that this approach will allow for the subsequent preparation of monoclonal antibodies of particular value from the reqmsite organ of the animal successfully resistmg or clearing infection for the development of prophylactic agents or therapeutic treatments of bactenal infection, particularly Staphylococcus aureus infection, m mammals, particularly humans The polypeptide may be used as an antigen for vaccmation of a host to produce specific antibodies which protect agamst mvasion of bactena, for example by blockmg adherence of bactena to damaged tissue Examples of tissue damage mclude wounds m sk n or connective tissue caused, e g , by mechanical, chemical or thermal damage or by implantation of indwelling devices, or wounds m the mucous membranes, such as the mouth, mammary glands, urethra or vagma The mvention also mcludes a vaccme formulation which compnses an immunogenic recombinant protem of the mvention together with a smtable earner Smce the protem may be broken down m the stomach, it is preferably administered parenterally, mcludmg, for example, administration that is subcutaneous, intramuscular, intravenous, or intradermal. Formulations suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxidants, buffers, bacteriostats and solutes which render the formulation insotonic with the bodily fluid, preferably the blood, of the individual; 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 ampules 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. While the invention has been described with reference to certain Histidine Kinase protein, it is to be understood that this covers fragments of the naturally occurring protein and similar proteins with additions, deletions or substitutions which do not substantially affect the immunogenic properties of the recombinant protein. Compositions, kits and administration The invention also relates to compositions comprising the polynucleotide or the polypeptides discussed above or their agonists or antagonists. The polypeptides of the invention may be employed in combination with a non-sterile or sterile carrier or carriers for use with cells, tissues or organisms, such as a pharmaceutical carrier suitable for administration to a subject. Such compositions comprise, for instance, a media additive or a therapeutically effective amount of a polypeptide of the invention and a pharmaceutically acceptable carrier or excipient. Such carriers may include, but are not limited to, saline, buffered saline, dextrose, water, glycerol, ethanol and combinations thereof. The formulation should suit the mode of aαrninistration. The invention further relates to diagnostic and pharmaceutical packs and kits comprising one or more containers filled with one or more of the ingredients of the aforementioned compositions of the invention. Polypeptides and other compounds of the invention may be employed alone or in conjunction with other compounds, such as therapeutic compounds. The pharmaceutical compositions may be administered in any effective, convenient manner including, for instance, administration by topical, oral, anal, vaginal, intravenous, intraperitoneal, intramuscular, subcutaneous, intranasal or intradermal routes among others. In therapy or as a prophylactic, the active agent may be administered to an mdividual as an lnjectable composition, for example as a stenle aqueous dispersion, preferably isotomc Alternatively the composition may be formulated for topical application for example m the form of ointments, creams, lotions, eye ointments, eye drops, ear drops. mouthwash, impregnated dressmgs and sutures and aerosols, and may contain appropnate conventional additives, mcludmg, for example, preservatives, solvents to assist drug penetration, and emollients m omtments and creams Such topical formulations may also contain compatible conventional earners, for example cream or ointment bases, and ethanol or oleyl alcohol for lotions Such earners may constitute from about 1% to about 98% by weight of the formulation, more usually they will constitute up to about 80% by weight of the formulation For administration to mammals, and particularly humans, it is expected that the daily dosage level of the active agent will be from 0 01 mg/kg to 10 mg/kg, typically around 1 mg/kg The physician m any event will determine the actual dosage which will be most suitable for an mdividual and will vary with the age, weight and response of the particular mdividual The above dosages are exemplary of the average case There can, of course, be mdividual instances where higher or lower dosage ranges are mented, and such are within the scope of this mvention In-dwelling devices mclude surgical implants, prosthetic devices and catheters, I e , devices that are mtroduced to the body of an mdividual and remam m position for an extended time Such devices mclude, for example, artificial jomts, heart valves, pacemakers, vascular grafts, vascular catheters, cerebrospmal fluid shunts, urinary catheters, contmuous ambulatory pentoneal dialysis (CAPD) catheters The composition of the mvention may be administered by injection to achieve a systemic effect against relevant bactena shortly before msertion of an m-dwellmg device Treatment may be contmued after surgery dunng the m-body time of the device In addition, the composition could also be used to broaden penoperative cover for any surgical technique to prevent bactenal wound infections, especially Staphylococcus aureus wound infections Many orthopaedic surgeons consider that humans with prosthetic jomts should be considered for antibiotic prophylaxis before dental treatment that could produce a bacteremia Late deep infection is a senous complication sometimes leadmg to loss of the prosthetic jomt and is accompanied by significant morbidity and mortality It may therefore be possible to extend the use of the active agent as a replacement for prophylactic antibiotics m this situation In addition to the therapy described above, the compositions of this mvention may be used generally as a wound treatment agent to prevent adhesion of bactena to matnx protems exposed in wound tissue and for prophylactic use m dental treatment as an alternative to, or m conjunction with, antibiotic prophylaxis Alternatively, the composition of the mvention may be used to bathe an indwelling device immediately before insertion The active agent will preferably be present at a concentration of lμg/ml to lOmg/ml for bathmg of wounds or indwelling devices A vaccme composition is convemently m mjectable form Conventional adjuvants may be employed to enhance the immune response A smtable umt dose for vaccmation is 0 5-5 microgram/kg of antigen, and such dose is preferably admimstered 1-3 tunes and with an interval of 1-3 weeks With the mdicated dose range, no adverse toxicological effects will be observed with the compounds of the mvention which would preclude their administration to suitable individuals Each reference disclosed herem is incorporated by reference herem m its entirety Any patent application to which this application claims pnonty is also incorporated by reference herem m its entirety GLOSSARY The following definitions are provided to facilitate understanding of certain terms used frequently herem "Dιsease(s)" means and disease caused by or related to infection by a bactena, mcludmg disease, such as, infections of the upper respiratory tract (e g , otitis media, bactenal tracheitis, acute epiglottitis, thyroiditis), lower respiratory (e g , empyema, lung abscess), cardiac (eg , infective endocarditis), gastrointestinal (e g , secretory diaπhoea, splemc absces, rettopentoneal abscess), CNS (e g , cerebral abscess), eye (e g , blephantis, conjunctivitis, keratitis, endophthalmitis, preseptal and orbital cellulitis, darcryocystitis), kidney and urinary tract (e g , epididymitis, intrarenal and pennephnc absces, toxic shock syndrome), skin (e g , impetigo, folhcuhtis, cutaneous abscesses, cellulitis, wound infection, bactenal myositis) bone and jomt (e g , septic arthntis, osteomyelitis) "Host cell" is a cell which has been transformed or transfected, or is capable of transformation or transfection by an exogenous polynucleotide sequence "Identity," as known m the art, is a relationship between two or more polypeptide sequences or two or more polynucleotide sequences, as determmed by comparing the sequences In the art. "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 "similanty" can be readily calculated by known methods, mcludmg but not limited to those descnbed m (Computational Molecular Biology, Lesk, A M . ed , Oxford University Press, New York, 1988, Bwcomputing Informatics and Genome Projects, Smith, D W , ed , Academic Press, New York, 1993, Computer Analysis of Sequence Data, Part I, Gnffin, A M , and Gnffin, H G , eds , Humana Press, New Jersey, 1994, Sequence Analysis in Molecular Biology, von Hemje, G , Academic Press, 1987, and Sequence Analysis Primer, Gnbskov, M and Devereux, J , eds , M Stockton Press, New York, 1991, and Canllo, H , and Lipman, D , SIAM J Applied Math , 48 1073 (1988) Prefeπed methods to determme identity are designed to give the largest match between the sequences tested Methods to determme identity and similanty are codified m publicly available computer programs Preferred computer program methods to determme identity and similanty between two sequences mclude, 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 , J Molec Biol 215 403-410 (1990) The BLAST X program is publicly available from NCBI and other sources (BLAST Manual, Altschul, S , et al , NCBI NLM NIH Bethesda, MD 20894, Altschul, S , et al , J Mol Biol 215 403-410 (1990) The well known Smith Waterman algonthm may also be used to determine identity Prefeπed embodiments mclude an isolated polynucleotide compnsmg a polynucleotide havmg at least a 50.60, 70, 80, 85, 90, 95, 97 or 100% identity to a polynucleotide reference sequence of SEQ ID NO 1, wherem said reference sequence may be identical to the sequence of SEQ ID NO 1 or may mclude up to a certam mteger number of nucleotide alterations as compared to the reference sequence, wherem said alterations are selected from the group consistmg of at least one nucleotide deletion, substitution or msertion, and wherem said alterations may occur at the 5' or 3' terminal positions of the reference nucleotide sequence or anywhere between those terminal positions, mterspersed either individually among the nucleotides m the reference sequence or m one or more contiguous groups within the reference sequence, and wherem said number of nucleotide alterations is determmed by multiplying the total number of nucleotides m SEQ ID NO 1 by the numerical percent of the respective percent identity and subtractmg that product from said total number of nucleotides in SEQ ID NO 1, or n < x - (x • y), wherem n is the number of nucleotide alterations, x is the total number of nucleotides m SEQ ID NO 1, and 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 wherem any non-mteger product of x and y is rounded down to the nearest mteger pnor to subtractmg it from x "Isolated" means altered "by the hand of man" from its natural state, i e , if it occurs m nature, it has been changed or removed from its onginal environment, or both For example, a polynucleotide or a polypeptide naturally present in a living organism is not "isolated," but the same polynucleotide or polypeptide separated from the coexisting matenals of its natural state is "isolated", as the term is employed herem Moreover, a polynucleotide or polypeptide that is mtroduced mto an orgamsm by transformation, genetic mampulation or by any other recombinant method is "isolated" even if it is still present m said organism, which orgamsm may be living or nonliving "Polynucleotide(s)" generally refers to any polynbonucleotide or polydeoxnbonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA "Polynucleotide(s)" mclude, without -imitation, smgle- and double-stranded DNA, DNA that is a mixture of smgle- and double- stranded regions or single-, double- and tnple-stranded regions, smgle- and double-stranded RNA, and RNA that is mixture of single- and double-stranded regions, hybnd molecules compnsmg DNA and RNA that may be single-stranded or, more typically, double-stranded, or tnple-stranded regions, or a mixture of single- and double-stranded regions In addition, "polynucleotide" as used herem refers to tnple-stranded regions compnsmg RNA or DNA or both RNA and DNA The sttands m such regions may be from the same molecule or from different molecules The regions may mclude all of one or more of the molecules, but more typically mvolve only a region of some of the molecules One of the molecules of a tnple-hehcal region often is an oligonucleotide As used herein, the term "polynucleotide(s)" also mcludes DNAs or RNAs as descnbed above that contain one or more modified bases Thus, DNAs or RNAs with backbones modified for stabihty or for other reasons are "polynucleotide(s)" as that term is intended herem Moreover, DNAs or RNAs compnsing unusual bases, such as inosine, or modified bases, such as tπtylated bases, to name just two examples, are polynucleotides as the term is used herem It will be appreciated that a great vanety of modifications have been made to DNA and RNA that serve many useful purposes known to those of skill m the art The term "polynucleotide(s)" as it is employed herem embraces such chemically, enzymatically or metabohcally modified forms of polynucleotides, as well as the chemical forms of DNA and RNA charactenstic of viruses and cells, mcludmg, for example, simple and complex cells "Polynucleotιde(s)" also embraces short polynucleotides often refeπed to as ohgonucleotide(s) "Polypeptide(s)" refers to any peptide or protem compnsmg two or more ammo acids joined to each other by peptide bonds or modified peptide bonds "Polypeptide(s)" refers to both short chains, commonly refeπed to as peptides, ohgopeptides and ohgomers and to longer chains generally refeπed to as proteins Polypeptides may contain ammo acids other than the 20 gene encoded ammo acids "Polypeptide(s)" mclude those modified either by natural processes, such as processmg and other post-ttanslahonal modifications, but also by chemical modification techniques Such modifications are well descnbed m basic texts and m more detailed monographs, as well as m a voluminous research literature, and they are well known to those of skill m the art It will be appreciated that the same type of modification may be present in the same or varying degree at several sites in a given polypeptide Also, a given polypeptide may contain many types of modifications Modifications can occur anywhere m a polypeptide, mcludmg the peptide backbone, the ammo acid side-chains, and the ammo or carboxyl termini Modifications mclude. for example, acetylation, acylation, ADP-nbosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide denvative, covalent attachment of a hpid or hpid denvative, covalent attachment of phosphotidyhnositol. cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cysteine, formation of pyroglutamate, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, lodination, methylation, mynstoylation, oxidation, proteolytic processmg, phosphorylation, prenylation, racemization, glycosylation, hpid attachment, sulfation, gamma- carboxylation of glutamic acid residues, hydroxylation and ADP-nbosylation, selenoylation, sulfation, transfer-RNA mediated addition of ammo acids to proteins, such as argmylation, and ubiquitination See, for instance, PROTEINS - STRUCTURE AND MOLECULAR PROPERTIES, 2nd Ed , T E Creighton, W H Freeman and Company, New York (1993) and Wold, F , Posttranslational Protem Modifications Perspectives and Prospects, pgs 1-12 in POSTTRANSLATIONAL COVALENT MODIFICATION OF PROTEINS, B C Johnson, Ed , Academic Press, New York (1983), Serfter et al , Meth Enzymol 182 626-646 (1990) and Rattan et al , Protein Synthesis Posttranslational Modifications and Aging, Ann N Y Acad Sci 663 48- 62 (1992) Polypeptides may be branched or cyclic, with or without branching Cyclic, branched and branched circular polypeptides may result from post-ttanslational natural processes and may be made by entirely synthetic methods, as well 'Nanant(s)" as the term is used herem, is a polynucleotide or polypeptide that differs from a reference polynucleotide or polypeptide respectively, but retams essential properties A typical vanant of a polynucleotide differs m nucleotide sequence from another, reference polynucleotide Changes in the nucleotide sequence of the vanant may or may not alter the ammo acid sequence of a polypeptide encoded by the reference polynucleotide Nucleotide changes may result m ammo acid substitutions, additions, deletions, fusions and truncations in the polypeptide encoded by the reference sequence, as discussed below A typical vanant 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 vanant are closely similar overall and, m many regions, identical A vanant and reference polypeptide may differ m ammo acid sequence by one or more substitutions, additions, deletions m any combination A substituted or inserted ammo 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 vanant, or it may be a vanant that is not known to occur naturally Non- naturally occurring vanants of polynucleotides and polypeptides may be made by mutagenesis techniques, by direct synthesis, and by other recombinant methods known to skilled artisans EXAMPLES The examples below are earned out usmg standard techniques, which are well known and routine to those of skill m the art, except where otherwise descnbed in detail The examples are illustrative, but do not limit the mvention Example 1 Strain selection, Library Production and Sequencing The polynucleotide havmg a DNA sequence given m Table 1 [SEQ ID NO 1] was obtained from a library of clones of chromosomal DNA of Staphylococcus aureus m E coli The sequencmg data from two or more clones containing overlappmg Staphylococcus aureus DNAs was used to construct the contiguous DNA sequence m SEQ ID NO 1 Libranes may be prepared by routme methods, for example Methods 1 and 2 below Total cellular DNA is isolated from Staphylococcus aureus WCUH 29 accordmg to standard procedures and size-fractionated by either of two methods Method 1 Total cellular DNA is mechanically sheared by passage through a needle m order to size-fractionate accordmg to standard procedures DNA fragments of up to 1 lkbp m size are rendered blunt by treatment with exonuclease and DNA polymerase, and EcoRI linkers added Fragments are ligated mto the vector Lambda ZapII that has been cut with EcoRI. the library packaged by standard procedures and E coli infected with the packaged library The library is amplified by standard procedures Method 2 Total cellular DNA is partially hydrolyzed with a one or a combmation of restriction enzymes appropriate to generate a senes of fragments for clonmg mto library vectors (e g . Rsal, Pall, Alul, Bshl235I), and such fragments are size-fractionated accordmg to standard procedures EcoRI linkers are ligated to the DNA and the fragments then ligated mto the vector Lambda ZapII that have been cut with EcoRI, the library packaged by standard procedures, and E coli infected with the packaged library The library is amplified by standard procedures

Claims: What is claimed is: 1 An isolated polynucleotide compnsmg a polynucleotide having at least a 70% identity to a polynucleotide encodmg a polypeptide compnsmg the ammo acid sequence of SEQ ID NO 2 2 An isolated polynucleotide compnsmg a polynucleotide having at least a 70% identity to a polynucleotide encodmg the same mature polypeptide expressed by the Histidme Kinase gene contained in the Staphylococcus aureus 3 An isolated polynucleotide compnsmg a polynucleotide encodmg a polypeptide compnsmg an ammo acid sequence which is at least 70% identical to the ammo acid sequence of SEQ ID NO 2 4 An isolated polynucleotide that is complementary to the polynucleotide of claim 1 5 The polynucleotide of Claim 1 wherem the polynucleotide is DNA or RNA 6 The polynucleotide of Claim 1 compnsmg the nucleic acid sequence set forth in SEQ ID NO l 7 The polynucleotide of Claim 1 compnsmg nucleotide 744 to the stop codon which begins at nucleotide number 2097 set forth SEQ ID NO 1 8 The polynucleotide of Claim 1 which encodes a polypeptide compnsmg the ammo acid sequence of SEQ ED NO 2 9 A vector compnsing the polynucleotide of Claim 1 10 A host cell compnsing the vector of Claim 9 11 A process for producmg a polypeptide compnsmg expressing from the host cell of Claim 10 a polypeptide encoded by said DNA 12 A process for producmg a Histidme Kinase polypeptide or fragment compnsmg cultunng a host of claim 10 under conditions sufficient for the production of said polypeptide or fragment 13 A polypeptide compnsing an ammo acid sequence which is at least 70% identical to the ammo acid sequence of SEQ ID NO 2 14 A polypeptide compnsmg an ammo acid sequence as set forth m SEQ ID NO 2 15 An antibody against the polypeptide of claim 14 16 An antagonist which inhibits the activity or expression of the polypeptide of claim 14 17 A method for the treatment of an mdividual in need of Histidme Kinase polypeptide compnsmg administering to the mdividual a therapeutically effective amount of the polypeptide of claim 14 18 A method for the treatment of an mdividual havmg need to inhibit Histidme Kinase polypeptide compnsmg administering to the mdividual a therapeutically effective amount of the antagonist of Claim 14 19 A process for diagnosing a disease related to expression or activity of the polypeptide of claim 14 in an mdividual compnsmg (a) determining a nucleic acid sequence encoding said polypeptide, and/or (b) analyzing for the presence or amount of said polypeptide m a sample denved from the mdividual 20 A method for identifying compounds which interact with and inhibit or activate an activity of the polypeptide of claim 14 compnsmg contacting a composition compnsmg the polypeptide with the compound to be screened under conditions to permit mteraction between the compoxmd and the polypeptide to assess the mteraction of a compound, such mteraction bemg associated with a second component capable of providing a detectable signal m response to the mteraction of the polypeptide with the compound, and determining whether the compound interacts with and activates or inhibits an activity of the polypeptide by detecting the presence or absence of a signal generated from the mteraction of the compound with the polypeptide 21 A method for mducmg an immimological response m a mammal which compnses inoculating the mammal with Histidme Kinase polypeptide of claim 14, or a fragment or vanant thereof, adequate to produce antibody and/or T cell immune response to protect said animal from disease 22 A method of mducmg immunological response m a mammal which compnses delivering a nucleic acid vector to direct expression of Histidme Kinase polypeptide of claim 14, or fragment or a vanant thereof, for expressmg said Histidme Kinase polypeptide, or a fragment or a vanant thereof in vivo m order to mduce an immunological response to produce antibody and/ or T cell immune response to protect said animal from disease 23 An isolated polynucleotide compnsing a polynucleotide havmg at least a 70% identity to a polynucleotide encodmg a polypeptide compnsmg the ammo acid sequence of SEQ ID NO 4 24 An isolated polynucleotide compnsmg a polynucleotide havmg at least a 70% identity to the polynucleotide sequence of SEQ ID NO 3

Inventors: DEBOUCK CHRISTINE, JAWORSKI DEBORAH D, MOONEY JEFFREY L, SHILLING LISA K, THROUP JOHN P, WALLIS NICOLA G, WANG MIN, ZHONG YIYI, DEBOUCK, CHRISTINE, JAWORSKI, DEBORAH, D., MOONEY, JEFFREY, L., SHILLING, LISA, K., THROUP, JOHN, P., WALLIS, NICOLA, G., WANG, MIN, ZHONG, YIYI
Applicants: SMITHKLINE BEECHAM CORP, SMITHKLINE BEECHAM CORPORATION
Application year: 1999
Main classification: C12N9
IPC codes: A61K38, A61K39, A61K45, A61P31, A61P37, C07H21, C07K14, C07K16, C12N1, C12N5, C12N9, C12N15, C12Q1, G01N33