1887

Abstract

The staphylococcal accessory regulator () plays a central role in the regulation of virulence in . To date, studies involving have focused on its activity as a global regulator that modulates transcription of a wide variety of genes (>100) and its role in virulence. However, there is also evidence to suggest the existence of accessory elements that modulate SarA production and/or function. A reporter system was developed to identify such elements, and a new gene, (SA1233), mutation of which results in reduced expression of SarA, was identified and characterized. Additionally, it was shown that mutation of resulted in altered transcription of the accessory gene regulator () and the genes encoding several virulence factors including alpha toxin () and protein A (). However, the impact of mutating was different in the laboratory strain RN6390 and the clinical isolate UAMS-1. For instance, mutation of caused a decrease in and transcription in RN6390 but had a different effect in UAMS-1. The strain-dependent effects of the mutation were similar to those observed previously, which suggests that may modulate the production of specific virulence factors through its impact on . Interestingly, sequence analysis of Msa suggests that it is a putative membrane protein with three membrane-spanning regions, indicating that Msa might interact with the environment. The findings show that is involved in the expression of SarA and several virulence factors.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.29071-0
2006-09-01
2019-10-24
Loading full text...

Full text loading...

/deliver/fulltext/micro/152/9/2559.html?itemId=/content/journal/micro/10.1099/mic.0.29071-0&mimeType=html&fmt=ahah

References

  1. Altschul, S. F., Madden, T. L., Schaffer, A. A., Zhang, J., Zhang, Z., Miller, W. & Lipman, D. J. ( 1997; ). Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res 25, 3389–3402.[CrossRef]
    [Google Scholar]
  2. Arvidson, S. & Tegmark, K. ( 2001; ). Regulation of virulence determinants in Staphylococcus aureus. Int J Med Microbiol 291, 159–170.[CrossRef]
    [Google Scholar]
  3. 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]
  4. Beenken, K. E., Blevins, J. S. & Smeltzer, M. S. ( 2003; ). Mutation of sarA in Staphylococcus aureus limits biofilm formation. Infect Immun 71, 4206–4211.[CrossRef]
    [Google Scholar]
  5. Bischoff, M., Entenza, J. M. & Giachino, P. ( 2001; ). Influence of a functional sigB operon on the global regulators sar and agr in Staphylococcus aureus. J Bacteriol 183, 5171–5179.[CrossRef]
    [Google Scholar]
  6. Blevins, J. S., Gillaspy, A. F., Rechtin, T. M., Hurlburt, B. K. & Smeltzer, M. S. ( 1999; ). The staphylococcal accessory regulator (sar) represses transcription of the Staphylococcus aureus collagen adhesin gene (cna) in an agr-independent manner. Mol Microbiol 33, 317–326.[CrossRef]
    [Google Scholar]
  7. Blevins, J. S., Beenken, K. E., Elasri, M. O., Hurlburt, B. K. & Smeltzer, M. S. ( 2002; ). Strain-dependent differences in the regulatory roles of sarA and agr in Staphylococcus aureus. Infect Immun 70, 470–480.[CrossRef]
    [Google Scholar]
  8. Blevins, J. S., Elasri, M. O., Allmendinger, S. D., Beenken, K. E., Skinner, R. A., Thomas, J. R. & Smeltzer, M. S. ( 2003; ). Role of sarA in the pathogenesis of Staphylococcus aureus musculoskeletal infection. Infect Immun 71, 516–523.[CrossRef]
    [Google Scholar]
  9. Booth, M. C., Cheung, A. L., Hatter, K. L., Jett, B. D., Callegan, M. C. & Gilmore, M. S. ( 1997; ). Staphylococcal accessory regulator (sar) in conjunction with agr contributes to Staphylococcus aureus virulence in endophthalmitis. Infect Immun 65, 1550–1556.
    [Google Scholar]
  10. Bronner, S., Monteil, H. & Prevost, G. ( 2004; ). Regulation of virulence determinants in Staphylococcus aureus: complexity and applications. FEMS Microbiol Rev 28, 183–200.[CrossRef]
    [Google Scholar]
  11. Cassat, J. E., Dunman, P. M., McAleese, F., Murphy, E., Projan, S. J. & Smeltzer, M. S. ( 2005; ). Comparative genomics of Staphylococcus aureus musculoskeletal isolates. J Bacteriol 187, 576–592.[CrossRef]
    [Google Scholar]
  12. Chakrabarti, S. K. & Misra, T. K. ( 2000; ). SarA represses agr operon expression in a purified in vitro Staphylococcus aureus transcription system. J Bacteriol 182, 5893–5897.[CrossRef]
    [Google Scholar]
  13. Chan, P. F. & Foster, S. J. ( 1998a; ). The role of environmental factors in the regulation of virulence-determinant expression in Staphylococcus aureus 8325-4. Microbiology 144, 2469–2479.[CrossRef]
    [Google Scholar]
  14. Chan, P. F. & Foster, S. J. ( 1998b; ). Role of SarA in virulence determinant production and environmental signal transduction in Staphylococcus aureus. J Bacteriol 180, 6232–6241.
    [Google Scholar]
  15. Cheung, A. L., Yeaman, M. R., Sullam, P. M., Witt, M. D. & Bayer, A. S. ( 1994; ). Role of the sar locus of Staphylococcus aureus in induction of endocarditis in rabbits. Infect Immun 62, 1719–1725.
    [Google Scholar]
  16. 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]
  17. Cheung, A. L., Bayer, A. S., Zhang, G., Gresham, H. & Xiong, Y. Q. ( 2004; ). Regulation of virulence determinants in vitro and in vivo in Staphylococcus aureus. FEMS Immunol Med Microbiol 40, 1–9.[CrossRef]
    [Google Scholar]
  18. Chien, Y., Manna, A. C. & Cheung, A. L. ( 1998; ). SarA level is a determinant of agr activation in Staphylococcus aureus. Mol Microbiol 30, 991–1001.[CrossRef]
    [Google Scholar]
  19. Chien, Y., Manna, A. C., Projan, S. J. & Cheung, A. L. ( 1999; ). SarA, a global regulator of virulence determinants in Staphylococcus aureus, binds to a conserved motif essential for sar-dependent gene regulation. J Biol Chem 274, 37169–37176.[CrossRef]
    [Google Scholar]
  20. Dunman, P. M., Murphy, E., Haney, S. & 7 other authors ( 2001; ). Transcription profiling-based identification of Staphylococcus aureus genes regulated by the agr and/or sarA loci. J Bacteriol 183, 7341–7353.[CrossRef]
    [Google Scholar]
  21. Francis, K. P., Joh, D., Bellinger-Kawahara, C., Hawkinson, M. J., Purchio, T. F. & Contag, P. R. ( 2000; ). Monitoring bioluminescent Staphylococcus aureus infections in living mice using a novel luxABCDE construct. Infect Immun 68, 3594–3600.[CrossRef]
    [Google Scholar]
  22. Gillaspy, A. F., Hickmon, S. G., Skinner, R. A., Thomas, J. R., Nelson, C. L. & Smeltzer, M. S. ( 1995; ). Role of the accessory gene regulator (agr) in pathogenesis of staphylococcal osteomyelitis. Infect Immun 63, 3373–3380.
    [Google Scholar]
  23. Gillaspy, A. F., Lee, C. Y., Sau, S., Cheung, A. L. & Smeltzer, M. S. ( 1998; ). Factors affecting the collagen binding capacity of Staphylococcus aureus. Infect Immun 66, 3170–3178.
    [Google Scholar]
  24. Goerke, C., Campana, S., Bayer, M. G., Doring, G., Botzenhart, K. & Wolz, C. ( 2000; ). Direct quantitative transcript analysis of the agr regulon of Staphylococcus aureus during human infection in comparison to the expression profile in vitro. Infect Immun 68, 1304–1311.[CrossRef]
    [Google Scholar]
  25. Heinrichs, J. H., Bayer, M. G. & Cheung, A. L. ( 1996; ). Characterization of the sar locus and its interaction with agr in Staphylococcus aureus. J Bacteriol 178, 418–423.
    [Google Scholar]
  26. Hirokawa, T., Boon-Chieng, S. & Mitaku, S. ( 1998; ). SOSUI: classification and secondary structure prediction system for membrane proteins. Bioinformatics 14, 378–379.[CrossRef]
    [Google Scholar]
  27. Horsburgh, M. J., Aish, J. L., White, I. J., Shaw, L., Lithgow, J. K. & Foster, S. J. ( 2002; ). σ B modulates virulence determinant expression and stress resistance: characterization of a functional rsbU strain derived from Staphylococcus aureus 8325-4. J Bacteriol 184, 5457–5467.[CrossRef]
    [Google Scholar]
  28. Janzon, L. & Arvidson, S. ( 1990; ). The role of the delta-lysin gene (hld) in the regulation of virulence genes by the accessory gene regulator (agr) in Staphylococcus aureus. EMBO J 9, 1391–1399.
    [Google Scholar]
  29. Jones, C. L. & Khan, S. A. ( 1986; ). Nucleotide sequence of the enterotoxin B gene from Staphylococcus aureus. J Bacteriol 166, 29–33.
    [Google Scholar]
  30. Karlsson, A., Saravia-Otten, P., Tegmark, K., Morfeldt, E. & Arvidson, S. ( 2001; ). Decreased amounts of cell wall-associated protein A and fibronectin-binding proteins in Staphylococcus aureus sarA mutants due to up-regulation of extracellular proteases. Infect Immun 69, 4742–4748.[CrossRef]
    [Google Scholar]
  31. Katzif, S., Danavall, D., Bowers, S., Balthazar, J. T. & Shafer, W. M. ( 2003; ). The major cold shock gene, cspA, is involved in the susceptibility of Staphylococcus aureus to an antimicrobial peptide of human cathepsin G. Infect Immun 71, 4304–4312.[CrossRef]
    [Google Scholar]
  32. Kullik, I., Giachino, P. & Fuchs, T. ( 1998; ). Deletion of the alternative sigma factor sigmaB in Staphylococcus aureus reveals its function as a global regulator of virulence genes. J Bacteriol 180, 4814–4820.
    [Google Scholar]
  33. Kyte, J. & Doolittle, R. F. ( 1982; ). A simple method for displaying the hydropathic character of a protein. J Mol Biol 157, 105–132.[CrossRef]
    [Google Scholar]
  34. Lindsay, J. A. & Foster, S. J. ( 1999; ). Interactive regulatory pathways control virulence determinant production and stability in response to environmental conditions in Staphylococcus aureus. Mol Gen Genet 262, 323–331.[CrossRef]
    [Google Scholar]
  35. Manna, A. C., Bayer, M. G. & Cheung, A. L. ( 1998; ). Transcriptional analysis of different promoters in the sar locus in Staphylococcus aureus. J Bacteriol 180, 3828–3836.
    [Google Scholar]
  36. Marchler-Bauer, A. & Bryant, S. H. ( 2004; ). CD-Search: protein domain annotations on the fly. Nucleic Acids Res 32, W327–W331.[CrossRef]
    [Google Scholar]
  37. McAleese, F. M., Walsh, E. J., Sieprawska, M., Potempa, J. & Foster, T. J. ( 2001; ). Loss of clumping factor B fibrinogen binding activity by Staphylococcus aureus involves cessation of transcription, shedding and cleavage by metalloprotease. J Biol Chem 276, 29969–29978.[CrossRef]
    [Google Scholar]
  38. McCallum, N., Bischoff, M., Maki, H., Wada, A. & Berger-Bachi, B. ( 2004; ). TcaR, a putative MarR-like regulator of sarS expression. J Bacteriol 186, 2966–2972.[CrossRef]
    [Google Scholar]
  39. McGavin, M. J., Zahradka, C., Rice, K. & Scott, J. E. ( 1997; ). Modification of the Staphylococcus aureus fibronectin binding phenotype by V8 protease. Infect Immun 65, 2621–2628.
    [Google Scholar]
  40. Morfeldt, E., Tegmark, K. & Arvidson, S. ( 1996; ). Transcriptional control of the agr-dependent virulence gene regulator, RNAIII, in Staphylococcus aureus. Mol Microbiol 21, 1227–1237.[CrossRef]
    [Google Scholar]
  41. Nilsson, I. M., Bremell, T., Ryden, C., Cheung, A. L. & Tarkowski, A. ( 1996; ). Role of the staphylococcal accessory gene regulator (sar) in septic arthritis. Infect Immun 64, 4438–4443.
    [Google Scholar]
  42. Novick, R. P. ( 1990; ). The staphylococcus as a molecular genetic system. In Molecular Biology of the Staphylococci, pp. 1–37. Edited by R. P. Novick. New York: VCH Publishers.
  43. Novick, R. P. ( 2003; ). Autoinduction and signal transduction in the regulation of staphylococcal virulence. Mol Microbiol 48, 1429–1449.[CrossRef]
    [Google Scholar]
  44. 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]
  45. Rechtin, T. M., Gillaspy, A. F., Schumacher, M. A., Brennan, R. G., Smeltzer, M. S. & Hurlburt, B. K. ( 1999; ). Characterization of the SarA virulence gene regulator of Staphylococcus aureus. Mol Microbiol 33, 307–316.[CrossRef]
    [Google Scholar]
  46. Rice, K. C., Patton, T., Yang, S. J., Dumoulin, A., Bischoff, M. & Bayles, K. W. ( 2004; ). Transcription of the Staphylococcus aureus cid and lrg murein hydrolase regulators is affected by sigma factor B. J Bacteriol 186, 3029–3037.[CrossRef]
    [Google Scholar]
  47. Rossi, J., Bischoff, M., Wada, A. & Berger-Bachi, B. ( 2003; ). MsrR, a putative cell envelope-associated element involved in Staphylococcus aureus sarA attenuation. Antimicrob Agents Chemother 47, 2558–2564.[CrossRef]
    [Google Scholar]
  48. Schmidt, K. A., Manna, A. C. & Cheung, A. L. ( 2003; ). SarT influences sarS expression in Staphylococcus aureus. Infect Immun 71, 5139–5148.[CrossRef]
    [Google Scholar]
  49. Sifri, C. D., Begun, J., Ausubel, F. M. & Calderwood, S. B. ( 2003; ). Caenorhabditis elegans as a model host for Staphylococcus aureus pathogenesis. Infect Immun 71, 2208–2217.[CrossRef]
    [Google Scholar]
  50. Smeltzer, M. S., Hart, M. E. & Iandolo, J. J. ( 1993; ). Phenotypic characterization of xpr, a global regulator of extracellular virulence factors in Staphylococcus aureus. Infect Immun 61, 919–925.
    [Google Scholar]
  51. Snodgrass, J. L., Mohamed, N., Ross, J. M., Sau, S., Lee, C. Y. & Smeltzer, M. S. ( 1999; ). Functional analysis of the Staphylococcus aureus collagen adhesin B domain. Infect Immun 67, 3952–3959.
    [Google Scholar]
  52. Somerville, G. A., Chaussee, M. S., Morgan, C. I., Fitzgerald, J. R., Dorward, D. W., Reitzer, L. J. & Musser, J. M. ( 2002; ). Staphylococcus aureus aconitase inactivation unexpectedly inhibits post-exponential-phase growth and enhances stationary-phase survival. Infect Immun 70, 6373–6382.[CrossRef]
    [Google Scholar]
  53. Sterba, K. M., Mackintosh, S. G., Blevins, J. S., Hurlburt, B. K. & Smeltzer, M. S. ( 2003; ). Characterization of Staphylococcus aureus SarA binding sites. J Bacteriol 185, 4410–4417.[CrossRef]
    [Google Scholar]
  54. Thompson, J. D., Gibson, T. J., Plewniak, F., Jeanmougin, F. & Higgins, D. G. ( 1997; ). The clustal_x windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25, 4876–4882.[CrossRef]
    [Google Scholar]
  55. Valle, J., Toledo-Arana, A., Berasain, C., Ghigo, J. M., Amorena, B., Penades, J. R. & Lasa, I. ( 2003; ). SarA and not sigmaB is essential for biofilm development by Staphylococcus aureus. Mol Microbiol 48, 1075–1087.[CrossRef]
    [Google Scholar]
  56. Wiltshire, M. D. & Foster, S. J. ( 2001; ). Identification and analysis of Staphylococcus aureus components expressed by a model system of growth in serum. Infect Immun 69, 5198–5202.[CrossRef]
    [Google Scholar]
  57. Wolz, C., Pohlmann-Dietze, P., Steinhuber, A., Chien, Y. T., Manna, A., van Wamel, W. & Cheung, A. ( 2000; ). Agr-independent regulation of fibronectin-binding protein(s) by the regulatory locus sar in Staphylococcus aureus. Mol Microbiol 36, 230–243.[CrossRef]
    [Google Scholar]
  58. Yamanaka, K. ( 1999; ). Cold shock response in Escherichia coli. J Mol Microbiol Biotechnol 1, 193–202.
    [Google Scholar]
  59. Yanisch-Perron, C., Vieira, J. & Messing, J. ( 1985; ). Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33, 103–119.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.29071-0
Loading
/content/journal/micro/10.1099/mic.0.29071-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error