GidA1

Details GidA1 GidA1

1998-06-24

2001-05-29

Navarro, Albert (Department: 1645)

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

C435S252300, C435S320100, C435S325000, C536S023700

Reexamination Certificate

active

06238882

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 gidA family, as well as their variants, hereinafter referred to as “gidA1,” “gidA1 polynucleotide(s),” and “gidA1 polypeptide(s)” as the case may be.
BACKGROUND OF THE INVENTION
The Streptococci make up a medically important genera of microbes known to cause several types of disease in humans, including, for example, otitis media, conjunctivitis, pneumonia, bacteremia, meningitis, sinusitis, pleural empyema and endocarditis, and most particularly meningitis, such as for example infection of cerebrospinal fluid. Since its isolation more than 100 years ago,
Streptococcus pneumoniae
has been one of the more intensively studied microbes. For example, much of our early understanding that DNA is, in fact, the genetic material was predicated on the work of Griffith and of Avery, Macleod and McCarty using this microbe. Despite the vast amount of research with
S. pneumoniae,
many questions concerning the virulence of this microbe remain. It is particularly preferred to employ Streptococcal genes and gene products as targets for the development of antibiotics.
The frequency of
Streptococcus pneumoniae
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
Streptococcus pneumoniae
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 gidA1 embodiments of the invention, that have a present benefit of, among other things, being useful to screen compounds for antimicrobial 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 gidA protein. The first described gidA gene was that of
E. coli
(von Meyenburg et al (1980) ICN-UCLA Symp. Mol. Cell. Biol. 19, 137-159; Genbank accession number P17112). The closest homolog of
S. pneumoniae
gidA1 is
Lactococcus lactis
gidA (Duwat, P. et al. (1997) J. Bacteriol. 179(14), 4473-79; Genbank accession number for polynucleotide is U80409, and SwissProt accession number for polypeptide is 032806). Other references relating to bacterial gidA genes are Ogasawara, N. & Yoshikawa, H. (1992) Mol. Microbiol. 6(5), 629-634, and Kunst, F. et al. (1997) Nature 390, 249-256 (
Bacillus subtilis
gidA); Karita, M et al. (1997) Infection & Immunity 65(10), 4158-64 (
Helicobacter pylori
gidA); Fsihi, H. et al. (1996) Microbiology 142(11), 3147-61 (
Mycobacterium leprae
gidA).
SUMMARY OF THE INVENTION
The present invention relates to gidA1, in particular gidA1 polypeptides and gidA1 polynucleotides, recombinant materials and methods of their production. In another aspect, the invention relates to methods for using such polypeptides and polynucleotides, including 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 agonist or antagonist 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 gidA1 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 gidA1 polypeptides and polynucleotides as described in greater detail below. In particular, the invention relates to polypeptides and polynucleotides of a gidA1 of
Streptococcus pneumoniae,
which is related by amino acid sequence homology to gidA polypeptide. The invention relates especially to gidA1 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
gidA1 Polynucleotide and Polypeptide Sequences
(A)
Streptococcus pneumoniae
gidA1 polynucleotide sequence [SEQ ID NO:1].

5′-

GAGGATATCCAGCTAGTTCCAGCCTTTTTAAAAACGGCCCTACCAGATTGGGAAGGCCAACTAAGACACATT

CATCTTGA

GGAATAGGAGAGAAACATGACTTATCATTTTACTGAAGAATACGATATTATTGTAATTGGTGCGGGACACGC

TGGGGTTG

AGGCTTCCTTGGCCGCTAGCCGTATGGGCTGTAAGGTCCTGCTTGCGACCATCAATATTGAAATGCTGGCTT

TCATGCCT

TGTAATCCCTCTATCGGTGGTTCTGCTAAGGGGATTGTCGTACGTGAAGTCGATGCCCTCGGTGGCGAGATG

GCCAAGAC

CATTGACAAGACTTACATCCAGATGAAGATGCTCAACACAGGGAAGGGCCCAGCCGTTCGTGCCCTTCGTGC

GCAGGCTG

ATAAGGAACTTTACTCTAAGGAAATGCGCAAGACAGTTGAAAATCAAGAAAATCTGACCCTTCGTCAAACCA

TGATTGAT

GAGATTTTGGTGGAAGATGGCAAGGTTGTCGGTGTGCGTACAGCCACCCATCAAGAATATGCTGCTAAGGCT

GTTATTGT

GACGACAGGGACTGCTCTCCGTGGGGAAATTATCATCGGAGACCTCAAGTACTCATCAGGTTCTAACCACAG

CTTGGCTT

CTATTAACCTAGCTGACAATCTCAAGGAACTGGGTCTCGAAATCGGTCGTTTCAAGACAGGAACCCCTCCAC

GTGTCAAG

GCTTCTTCTATCAATTACGATGTGACGGAAATTCAGCCAGGAGACGAAGTGCCTAATCATTTCTCATACACT

TCACGTGA

TGAGGATTATGTCAAAGATCAAGTGCCATGCTGGTTGACCTATACCAATGGTACCAGTCATGAGATTATCCA

AAACAACC

TCCACCGTGCGCCTATGTTTACAGGTGTGGTCAAGGGAGTGGGGCCTCGTTACTGTCCGTCGATTGAAGACA

AGATTGTG

CGCTTTGCGGACAAGGAACGTCACCAACTCTTCCTTGAGCCAGAAGGACGCAATACTGAGGAAGTCTATGTT

CAAGGACT

TTCAACCAGTCTGCCTGAGGATGTCCAGCGTGACTTGGTTCATTCCATCAAAGGTTTGGAAAATGCAGAGAT

GATGCGGA

CAGGTTATGCTATTGAGTATGATATGGTCTTGCCTCATCAGTTGCGTGCGACTTTGGAAACCAAGAAAATCT

CAGGTCTC

TTCACTGCTGGTCAGACAAATGGAACATCAGGTTATGAAGAAGCTGCTGGCCAAGGGATTATCGCGGGTATC

AATGCGGC

TCTGAAAATCCAAGGTAAACCTGAGTTGATTCTAAAACGAAGTGACGGTTATATCGGGGTGATGATCGACGA

CTTGGTGA

CCAAGGGAACCATTGAACCTTACCGTCTCTTGACCAGTCGTGCTGAATACCGTCTCATTCTTCGTCATGACA

ATGCTGAT

ATGCGCTTGACTGAGATGGGACGCGAGATTGGCCTTGTGGATGATGAACGCTGGGCTCGTTTTGAAATCAAG

AAAAATCA

ATTTGATAATGAGATGAAACGCCTAGACAGTATCAAACTCAAGCCAGTCAAGGAAACCAATGCTAAGGTTGA

GGAAATGG

GCTTCAAGCCGTTGACAGATGCGGTGACAGCCAAAGAATTCCTTCGCCGTCCAGAAGTTTCTTACCAAGATG

TGGTGGCC

TTCATCGGACCAGCTGCAGAAGACTTGGATGACAAGATTATCGAATTGATTGAAACAGAAATCAAGTACGAA

GGCTATAT

TTCAAAAGCCATGGATCAGGTTGCCAAGATGAAACGTATGGAAGAAAAACGCATTCCAGCCAATATTGACTG

GGATGACA

TCGATTCTATTGCGACGGAAGCTCGTCAGAAGTTCAAACTCATCAATCCAGAAACCATCGGCCAAGCCAGCC

GTATTTCG

GGAGTAAACCCAGCAGATATTTCTATTTTGATGGTGTATCTGGAAGGTAAAAATCGTAGTATTTCTAAAACT

CTTCAAAA

ATCAAAATGATACGTCGTCGGCTTCTTACGAATGAGTTCAAAGCTTGGCTTTGATTCATCTCCAGCCTCCCA

TAGTTCCC

CGAACTATGGGAGCTAACTC-3′

(B)
Streptococcus pneumoniae
gidA1 polypeptide sequence deduced
from a polynucleotide sequence in thi

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