1887

Abstract

A gene originally identified by signature-tagged mutagenesis as being required for virulence was cloned, sequenced and named . Hydropathy profiles revealed that SvrA is likely to be membrane associated, having two regions with six membrane-spanning domains, the regions separated by an extended hydrophilic loop. When compared with the wild-type strain, an mutant expressed greatly reduced amounts of α-, β- and δ-toxins and an increased amount of protein A. Toxin production by the mutant strain was restored to wild-type levels when complemented with a plasmid expressing the gene. Northern hybridization with probes specific for (encoding α-toxin) and (encoding protein A) showed that the mutant strain was affected in the transcription of these genes. mRNA was present in wild-type and strains, but mRNA and RNAIII were absent in the mutant strain. Virulence studies suggested that the attenuation of the mutant was probably due to its direct or indirect effect on the regulon. These results indicate that is required for the expression of and RNAIII transcripts and is therefore a new component of the regulatory network controlling virulence gene expression in .

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2002-10-01
2024-12-14
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References

  1. Abdelnour A, Arvidson S, Bremell T, Ryden C., Tarkowski A. 1993; The accessory gene regulator ( agr ) controls Staphylococcus aureus virulence in a murine arthritis model. Infect Immun 61:3879–3885
    [Google Scholar]
  2. Balaban N., Novick R. P. 1995; Autocrine regulation of toxin synthesis by Staphylococcus aureus . Proc Natl Acad Sci USA 92:1619–1623
    [Google Scholar]
  3. Balaban N, Goldkorn T, Nhan R. T. 8 other authors 1998; Autoinducer of virulence as a target for vaccine and therapy against Staphylococcus aureus . Science 280:438–440
    [Google Scholar]
  4. Balaban N, Goldkorn T, Gov Y, Hirshberg M, Koyfman N, Matthews H. R, Nhan R. T, Singh B., Uziel O. 2001; Regulation of Staphylococcus aureus pathogenesis via target of RNAIII-activating Protein (TRAP). J Biol Chem 276:2658–2667
    [Google Scholar]
  5. Bayer M. G, Heinrichs J. H., Cheung A. L. 1996; The molecular architecture of the sar locus in Staphylococcus aureus . J Bacteriol 178:4563–4570
    [Google Scholar]
  6. Beuzón C. R., Holden D. W. 2001; Use of mixed infections with Salmonella strains to study virulence genes and their interactions in vivo . Microbes Infect 3:1345–1352
    [Google Scholar]
  7. Bramley A. J, Patel A. H, O’Reilly M, Foster R., Foster T. J. 1989; Roles of alpha-toxin and beta-toxin in virulence of Staphylococcus aureus for the mouse mammary gland. Infect Immun 57:2489–2494
    [Google Scholar]
  8. Bunce C, Wheeler L, Reed G, Musser J., Barg N. 1992; Murine model of cutaneous infection with gram-positive cocci. Infect Immun 60:2636–2640
    [Google Scholar]
  9. Chan P. F., Foster S. J. 1998; Role of SarA in virulence determinant production and environmental signal transduction in Staphylococcus aureus . J Bacteriol 180:6232–6241
    [Google Scholar]
  10. Cheung A. L., Ying P. 1994; Regulation of alpha- and beta-hemolysins by the sar locus of Staphylococcus aureus . J Bacteriol 176:580–585
    [Google Scholar]
  11. Cheung A. L, Koomey J. M, Butler C. A, Projan S. J., Fischetti V. A. 1992; Regulation of exoprotein expression in Staphylococcus aureus by a locus ( sar ) distinct from agr . Proc Natl Acad Sci USA 89:6462–6466
    [Google Scholar]
  12. Cheung A. L, Eberhardt K. J, Chung E, Yeaman M. R, Sullam P. M, Ramos M., Bayer A. S. 1994; Diminished virulence of a sar -/ agr - mutant of Staphylococcus aureus in the rabbit model of endocarditis. J Clin Investig 94:1815–1822
    [Google Scholar]
  13. Cheung A. L, Bayer M. G., Heinrichs J. H. 1997; sar genetic determinants necessary for transcription of RNAII and RNAIII in the agr locus of Staphylococcus aureus . J Bacteriol 179:3963–3971
    [Google Scholar]
  14. Fournier B., Hooper D. C. 2000; A new two-component regulatory system involved in adhesion, autolysis, and extracellular proteolytic activity of Staphylococcus aureus . J Bacteriol 182:3955–3964
    [Google Scholar]
  15. Giraudo A. T, Cheung A. L., Nagel R. 1997; The sae locus of Staphylococcus aureus controls exoprotein synthesis at the transcriptional level. Arch Microbiol 168:53–58
    [Google Scholar]
  16. Hofmann K., Stoffel W. 1993; TMbase – a database of membrane spanning proteins segments. Biol Chem 374:166
    [Google Scholar]
  17. Ji G, Beavis R. C., Novick R. P. 1995; Cell density control of staphylococcal virulence mediated by an octapeptide pheromone. Proc Natl Acad Sci USA 92:12055–12059
    [Google Scholar]
  18. Ji G, Beavis R. C., Novick R. P. 1997; Bacterial interference caused by autoinducing peptide variants. Science 276:2027–2030
    [Google Scholar]
  19. Kayser F. H, Wust J., Corrodi P. 1973; Transduction and elimination of resistance determinants in methicillin-resistant Staphylococcus aureus . Antimicrob Agents Chemother 2:217–223
    [Google Scholar]
  20. Kornblum J, Kreiswirth B. N, Projan S. J, Ross H., Novick R. P. 1990; Agr: a polycistronic locus regulating exoprotein synthesis. In Molecular Biology of the Staphylococci pp 373–402 Edited by Novick R. P. New York: VCH Publishers;
    [Google Scholar]
  21. Kuroda M, Ohta T, Uchiyama I. 34 other authors 2001; Whole genome sequencing of methicillin-resistant Staphylococcus aureus . Lancet 357:1225–1240
    [Google Scholar]
  22. Lina G, Jarraud S, Ji G, Greenland T, Pedraza A, Etienne J, Novick R. P., Vandenesch F. 1998; Transmembrane topology and histidine protein kinase activity of AgrC, the agr signal receptor in Staphylococcus aureus . Mol Microbiol 28:655–662
    [Google Scholar]
  23. Lyngso R. B, Zuker M., Pedersen C. N. 1999; Fast evaluation of internal loops in RNA secondary structure prediction. Bioinformatics 15:440–445
    [Google Scholar]
  24. Macrina F. L, Wood P. H., Jones K. R. 1980; Genetic transformation of Streptococcus sanguis (Challis) with cryptic plasmids from Streptococcus ferus . Infect Immun 28:692–699
    [Google Scholar]
  25. McNamara P. J, Milligan-Monroe K. C, Khalili S., Proctor R. A. 2000; Identification, cloning, and initial characterization of rot, a locus encoding a regulator of virulence factor expression in Staphylococcus aureus . J Bacteriol 182:3197–3203
    [Google Scholar]
  26. Mei J. M, Nourbakhsh F, Ford C. W., Holden D. W. 1997; Identification of Staphylococcus aureus virulence genes in a murine model of bacteraemia using signature-tagged mutagenesis. Mol Microbiol 26:399–407
    [Google Scholar]
  27. Morfeldt E, Panova-Sapundjieva I, Gustafsson B., Arvidson S. 1996; Detection of the response regulator AgrA in the cytosolic fraction of Staphylococcus aureus by monoclonal antibodies. FEMS Microbiol Lett 143:195–201
    [Google Scholar]
  28. Novick R. P, Ross H. F, Projan S. J, Kornblum J, Kreiswirth B., Moghazeh S. 1993; Synthesis of staphylococcal virulence factors is controlled by a regulatory RNA molecule. EMBO J 12:3967–3975
    [Google Scholar]
  29. Novick R. P, Projan S. J, Kornblum J, Ross H. F, Ji G, Kreiswirth B, Vandenesch F., Moghazeh S. 1995; The agr P2 operon: an autocatalytic sensory transduction system in Staphylococcus aureus . Mol Gen Genet 248:446–458
    [Google Scholar]
  30. Peng H. L, Novick R. P, Kreiswirth B, Kornblum J., Schlievert P. 1988; Cloning, characterization, and sequencing of an accessory gene regulator ( agr ) in Staphylococcus aureus . J Bacteriol 170:4365–4372
    [Google Scholar]
  31. Pospiech A., Neumann B. 1995; A versatile quick-prep of genomic DNA from gram-positive bacteria. Trends Genet 11:217–218
    [Google Scholar]
  32. Qazi S. N, Counil E, Morrissey J, Rees C. E, Cockayne A, Winzer K, Chan W. C, Williams P., Hill P. J. 2001; agr expression precedes escape of internalized Staphylococcus aureus from the host endosome. Infect Immun 69:7074–7082
    [Google Scholar]
  33. Sambrook J, Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  34. Sherris J. C., Plorde J. J. 1990; Staphylococci. In Medical Microbiology: an Introduction to Infectious Diseases pp 275–289 Edited by Sherris J. C. New York: Elsevier;
    [Google Scholar]
  35. Wilson C. R, Skinner S. E., Shaw W. V. 1981; Analysis of two chloramphenicol resistance plasmids from Staphylococcus aureus : insertional inactivation of Cm resistance, mapping of restriction sites, and construction of cloning vehicles. Plasmid 5:245–258
    [Google Scholar]
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