Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues
Reexamination Certificate
2000-08-31
2002-03-05
Duffy, Patricia A. (Department: 1645)
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
C424S185100, C424S190100, C424S192100, C424S234100
Reexamination Certificate
active
06353093
ABSTRACT:
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 polynucleotides and polypeptides of the gidB family, as well as their variants, hereinafter referred to as “gidB,” “gidB polynucleotide(s),” and “gidB polypeptide(s)” as the case may be.
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.
S. aureus
is the second leading cause of bacteremia in cancer patients. Osteomyelitis, septic arthritis, septic thrombophlebitis and acute bacterial 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 risen 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 an unmet medical need and demand for new anti-microbial agents, vaccines, drug screening methods, and diagnostic tests for this organism.
Moreover, the drug discovery process is currently undergoing a fundamental revolution as it embraces “functional genomics,” that is, high throughput genome- or gene-based biology. This approach is rapidly superseding earlier approaches based on “positional cloning” and other methods. Functional genomics relies heavily on the various tools of bioinformatics to identify gene sequences of potential interest from the many molecular biology databases now available as well as from other sources. There is a continuing and significant need to identify and characterize further genes and other polynucleotides sequences and their related polypeptides, as targets for drug discovery.
Clearly, there exists a need for polynucleotides and polypeptides, such as the gidB embodiments of the invention, that have a present benefit of, among other things, being 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 characterization 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 significant amino acid sequence homology to a known gidB protein.
SUMMARY OF THE INVENTION
The present invention relates to gidB, in particular gidB polypeptides and gidB polynucleotides, recombinant materials and methods for their production. In another aspect, the invention relates to methods for using such polypeptides and polynucleotides, including the treatment of microbial diseases, amongst others. In a further aspect, the invention relates to methods for identifying agonists and antagonists using the materials provided by the invention, and for treating microbial infections and conditions associated with such infections with the identified compounds. In a still further aspect, the invention relates to diagnostic assays for detecting diseases associated with microbial infections and conditions associated with such infections, such as assays for detecting gidB expression or activity.
Various changes and modifications within the spirit and scope of the disclosed invention will become readily apparent to those skilled in the art from reading the following descriptions and from reading the other parts of the present disclosure.
DESCRIPTION OF THE INVENTION
The invention relates to gidB polypeptides and polynucleotides as described in greater detail below. In particular, the invention relates to polypeptides and polynucleotides of a gidB of
Staphylococcus aureus,
which is related by amino acid sequence homology to gidB polypeptide. The invention relates especially to gidB having the nucleotide and amino acid sequences set out in Table 1 as SEQ ID NO: 1 or 3 and SEQ ID NO: 2 or 4 respectively.
TABLE 1
gidB Polynucleotide and Polypeptide Sequences
(A)
Staphylococcus aureus
gidB polynucleotide sequence [SEQ ID NO:1].
5′-
TCTATATTATTGATTTACTTAGAACAAGGTAAACTCCAAAGGGTGAGTGACTAATGACTGTAGAATGGTTAG
CAGAACAA
TTAAAAGAACATAATATTGAATTAACTGAGACTCAAAAACAACAGTTTCAAACATATTATCGTTTACTTGTT
GAATGGAA
TGAAAAGATGAATTTGACAAGTATTACAGATGAACACGATGTATATTTGAAACATTTTTATGATTCCATTGC
ACCTAGTT
TTTATTTTGATTTTAATCAGCCTATAAGTATATGTGATGTAGGCGCTGGAGCTGGTTTTCCAAGTATTCCGT
TAAAAATA
ATGTTTCCGCAGTTAAAAGTGACGATTGTTGATTCATTAAATAAGCGTATTCAATTTTTAAACCATTTAGCG
TCAGAATT
ACAATTACAGGATGTCAGCTTTATACACGATAGAGCAGAAACATTTGGTAAGGGTGTCTACAGGGAGTCTTA
TGATGTTG
TTACTGCAAGAGCAGTAGCTAGATTATCCGTGTTGAGTGAATTGTGTTTACCGCTAATTAAAAAAGGTGGAC
AGTTTGTT
GCATTAAAATCTTCAAAAGGTGAAGAAGAATTAGAAGAAGCAAAATTTGCAATTAGTGTGTTAGGTGGTAAC
GTTACAGA
AACACATACCTTTAAATTGCCAGAAGATGCTGGAGAGCGCCAGATGTTCATTATTGATAAAAAAAGACAGAC
GCCGAAAA
AGTACCCAAGAAAACCAGGGACGCCTAATAAGACTCCTTTACTTGAAAAATAATGCATAATCCTTTACAACT
AACATAAA
AGGAGCGAAT-3′
(B)
Staphylococcus aureus
gidB polypeptide sequence deduced from a polynucleotide
sequence in this table [SEQ ID NO:2].
NH
2
-
MTVEWLAEQLKEHNIELTETQKQQFQTYYRLLVEWNEKMNLTSITDEHDVYLKHFYDSIAPSFYFDFNQPIS
ICDVGAGA
GFPSIPLKIMFPQLKVTIVDSLNKRIQFLNHLASELQLQDVSFIHDRAETFGKGVYRESYDVVTARAVARLS
VLSELCLP
LIKKGGQFVALKSSKGEEELEEAKFAISVLGGNVTETHTFKLPEDAGERQMFIIDKKRQTPKKYPRKPGTPN
KTPLLEK-COOH
(C)
Staphylococcus aureus
gidB ORF sequence [SEQ ID NO:3].
5′-
GTAAATCCAGCAGACATATCTATATTATTGATTTACTTAGAACAAGGTAAACTCCAAAGGGTGAGTGACTAA
TGACTGTA
GAATGGTTAGCAGAACAATTAAAAGAACATAATATTGAATTAACTGAGACTCAAAAACAACAGTTTCAAACA
TATTATCG
TTTACTTGTTGAATGGAATGAAAAGATGAATTTGACAAGTATTACAGATGAACACGATGTATATTTGAAACA
TTTTTATG
ATTCCATTGCACCTAGTTTTTATTTTGATTTTAATCAGCCTATAAGTATATGTGATGTAGGCGCTGGAGCTG
GTTTTCCA
AGTATTCCGTTAAAAATAATGTTTCCGCAGTTAAAAGTGACGATTGTTGATTCATTAAATAAGCGTATTCAA
TTTTTAAA
CCATTTAGCGTCAGAATTACAATTACAGGATGTCAGCTTTATACACGATAGAGCAAAAACATNTGGTAAGGG
TGTCTACA
NGGAGTCTTATGATGTTGTTACTGCAAGAGCAGTANCTAAATTATCCGTGTTGAGTGAATTGTGTTTACCGC
TAATTAAA
AAAGGTGGACAGTNTGTTGCATTAAAATCTTCAAAAGGTGAAGAAAAATTANAAAAAACANAATTTGCAATT
AGTGTGTT
AGGTGGTAACGTTACAGAAACACATACCTTTAAATTGCCAGAAGATGCTGGAGAGCGCCAGATGTTCATTAT
TGATAAAA
AAAGACAGACGCCGAAAAAGTACCCAAGAAAACCAGGGACGCCTAATAAGACTCCTTTACTTGAAAAATAAT
GCATAATC
CTTTACAACTAACATAAAAGGAGCGAATGGATAATGAAAAAACCTTTTTCAAAATTATTTGGTTTGAAAAAC
AAAGATGA
CATCATTGGACATATTGAAG-3′
(D)
Staphylococcus aureus
gidB polypeptide sequence deduced from a polynucleotide ORF
sequence in this table
[SEQ ID NO:4].
NH
2
-
MTVEWLAEQLKEHNIELTETQKQQFQTYYRLLVEWNEKMNLTSITDEHDVYLKRFYDSIAPSFYFDFNQPIS
ICDVGAGA
GFPSIPLKIMFPQLKVTIVDSLNKRIQFLNHLASELQLQDVSFIHDRAKTXGKGVYXESYDVVTARAVXKLS
VLSELCLP
LIKKGGQXVALKSSKGEEKLXKTXFAISVLGGNVTETHTFKLPEDAGERQMFIIDKKRQTPKKYPRKPGTPN
KTPLLEK-COOH
Deposited materials
A deposit containing a
Staphylococcus aureus
WCUH 29 strain has been deposited with the National Collections of Industrial and Marine Bacteria Ltd. (herein “NCIMB”), 23 St. Machar Drive, Aberdeen AB2 1RY, Scotland on Sep. 11, 1995 and assigned NCIMB Deposit No. 40771, and referred to as
Staphylococcus aureus
WCUH29 on deposit. The
Staphylococcus aureus
strain deposit is referred to herein as “the deposi
Burnham Martin K R
Kallender Howard
Lenox Anna Lisa
Ward Judith
Deibert Thomas S.
Duffy Patricia A.
Gimmi Edward R.
King William T.
SmithKline Beecham Corporation
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