Member of ATP dependent protease family, clpL

Details Member of ATP dependent protease family, clpL Member of ATP dependent protease family, clpL

1999-10-13

2002-09-10

Allen, Marianne P. (Department: 1631)

Drug, bio-affecting and body treating compositions

Enzyme or coenzyme containing

Hydrolases

C530S350000, C424S094640, C514S002600, C514S012200, C435S195000, C435S212000, C435S219000, C435S220000

Reexamination Certificate

active

06447773

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, in these and in other regards, the invention relates to novel polynucleotides and polypeptides of the ATP dependent proteases family, hereinafter referred to as “clpL”.
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 surface and deep tissues.
Staphylococcus 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 20 years. 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 has created a demand for both new anti-microbial agents and diagnostic tests for this organism
Recently several novel approaches have been described which purport to follow global gene expression during infection (Chuang, S. et al., (1993); Mahan, M. J. et al., Science 259:686-688 (1993); Hensel, M. et al., Science 269:400-403 (1995). These new techniques have so far been demonstrated with gram negative pathogen infections, but not with infections with gram positives presumably because the much slower development of global transposon mutagenesis and suitable vectors needed for these strategies in these organisms, and in the case of that process described by Chuang, S. et al., J. Bacteriol. 175:2026-2036 (1993), the difficulty of isolating suitable quantities of bacterial RNA free of mammalian RNA derived from the infected tissue to furnish bacterial RNA labelled to sufficiently high specific activity.
The present invention employs a novel technology to determine gene expression in the pathogen at different stages of infection of the mammalian host. A novel aspect of this invention is the use of a suitably labelled oligonucleotide probe which anneals specifically to the bacterial ribosomal RNA in Northern blots of bacterial RNA preparations from infected tissue. Using the more abundant ribosomal RNA as a hybridization target greatly facilitates the optimization of a protocol to purify bacterial RNA of a suitable size and quantity for RT-PCR from infected tissue.
A suitable oligonucleotide useful for applying this method to genes expressed in
Staphylococcus aureus
is 5′-GCTCCTAAAAGGTTACTCCACCGGC-3′ [SEQ ID NO:5].
Use of the technology of the present invention enables identification of bacterial genes transcribed during infection, inhibitors of which would have utility in anti-bacterial therapy. Specific inhibitors of such gene transcription or of the subsequent translation of the resultant mRNA or of the function of the corresponding expressed proteins would have utility in anti-bacterial therapy. This invention provides
Staphylococcus aureus
WCUH29 polynucleotides which are transcribed in infected tissue.
Clearly, there is a need for factors, such as the novel compounds of the invention, that have a present benefit of 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 which can play a role in preventing, ameliorating or correcting infections, dysfunctions or diseases.
The polypeptides of the invention have amino acid sequence homology to a known
Lactococcus lactis
ClpL protein (Genbank Accession Number: X62333).
SUMMARY OF THE INVENTION
It is an object of the invention to provide polypeptides that have been identified as novel clpL polypeptides by homology between the amino acid sequence set out in Table 1 [SEQ ID NO: 2] and a known amino acid sequence or sequences of other proteins such as Lactococcus lactis ClpL protein.
It is a further object of the invention to provide polynucleotides that encode clpL polypeptides, particularly polynucleotides that encode the polypeptide herein designated clpL.
In a particularly preferred embodiment of the invention, the polynucleotide comprises a region encoding clpL polypeptides comprising the sequence set out in Table 1 [SEQ ID NO: 1] which includes a fill length gene, or a variant thereof.
In another particularly preferred embodiment of the invention, there is a novel clpL protein from
Staphylococcus aureus
comprising the amino acid sequence of Table 1 [SEQ ID NO:2], or a variant thereof.
In accordance with another aspect of the invention, there is provided an isolated nucleic acid molecule encoding a mature polypeptide expressible by the
Staphylococcus aureus
WCUH 29 strain contained in the deposited stain.
As a further aspect of the invention, there are provided isolated nucleic acid molecules encoding clpL, particularly
Staphylococcus aureus
clpL, including mRNAs, cDNAs, genomic DNAs. Further embodiments of the invention include biologically, diagnostically, prophylactically, clinically or therapeutically useful variants thereof and compositions comprising the same.
In accordance with another aspect of the invention, there is provided the use of a polynucleotide of the invention for therapeutic or prophylactic purposes, in particular genetic immunization. Among the particularly preferred embodiments of the invention are naturally occurring allelic variants of clpL and polypeptides encoded thereby.
As another aspect of the invention, there are provided novel polypeptides of
Staphylococcus aureus
referred to herein as clpL as well as biologically, diagnostically, prophylactically, clinically or therapeutically useful variants thereof, and compositions comprising the same.
Among the particularly preferred embodiments of the invention are variants of clpL polypeptide encoded by naturally occurring alleles of the clpL gene.
In a preferred emnbodiment of the invention, there are provided methods for producing the aforementioned clpL polypeptides.
In accordance with yet another aspect of the invention, there are provided inhibitors to such polypeptides, useful as antibacterial agents, including, for example, antibodies.
In accordance with certain preferred embodiments of the invention, there are provided products, compositions and methods for assessing clpL expression, treating disease, for example, disease, such as, infections of the upper respiratory tract (e.g., otitis media, bacterial tracheitis, acute epiglottitis, thyroiditis), lower respiratory (e.g., empyema, lung abscess), cardiac (e.g., infective endocarditis), gastrointestinal (e.g., secretory diarrhoea, splenic abscess, retroperitoneal abscess), CNS (e.g., cerebral abscess), eye (e.g., blepharitis, conjunctivitis, keratitis, endophthalmitis, preseptal and orbital cellulitis, darcryocystitis), kidney and urinary tract (e.g., epididymitis, intrarenal and perinephric abscess, toxic shock syndrome), skin (e.g., impetigo, folliculitis, cutaneous abscesses, cellulitis, wound infection, bacterial myositis) bone and joint (e.g., septic arthritis, osteomyelitis), assaying genetic variation, and administering a clpL polypept

Affiliated with

Also associated with

Rating

Member of ATP dependent protease family, clpL does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Member of ATP dependent protease family, clpL, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Member of ATP dependent protease family, clpL will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-2822205