1887

Abstract

produces several virulence factors that allow it to cause a variety of infections. One of the major virulence factors is the capsule, which contributes to the survival of the pathogen within the host as a way to escape phagocytosis. The production of the capsular polysaccharide is encoded in a 16 gene operon, which is regulated in response to several environmental stimuli including nutrient availability. For instance, the capsule is produced in the late- and post-exponential growth phases, but not in the early- or mid-exponential growth phase. Several regulators are involved in capsule production, but the regulation of the operon is still poorly understood. In this study, we show that MsaB activates the operon by binding directly to a 10 bp repeat in the promoter region. We show that despite the fact that MsaB is expressed throughout four growth phases, it only activates capsule production in the late- and post-exponential growth phases. Furthermore, we find that MsaB does not bind to its target site in the early and mid-exponential growth phases. This correlates with decreased nutrient availability and capsule production. These data suggest either that MsaB binding ability changes in response to nutrients or that other operon regulators interfere with the binding of MsaB to its target site. This study increases our understanding of the regulation of capsule production and the mechanism of action of MsaB.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000243
2016-03-01
2024-04-16
Loading full text...

Full text loading...

/deliver/fulltext/micro/162/3/575.html?itemId=/content/journal/micro/10.1099/mic.0.000243&mimeType=html&fmt=ahah

References

  1. Arbeit R. D., Karakawa W. W., Vann W. F., Robbins J. B. 1984; Predominance of two newly described capsular polysaccharide types among clinical isolates of Staphylococcus aureus . Diagn Microbiol Infect Dis 2:85–91 [View Article][PubMed]
    [Google Scholar]
  2. Bae T., Schneewind O. 2006; Allelic replacement in Staphylococcus aureus with inducible counter-selection. Plasmid 55:58–63 [View Article][PubMed]
    [Google Scholar]
  3. Bagnoli F., Bertholet S., Grandi G. 2012; Inferring reasons for the failure of Staphylococcus aureus vaccines in clinical trials. Front Cell Infect Microbiol 2:16 [View Article][PubMed]
    [Google Scholar]
  4. Bischoff M., Dunman P., Kormanec J., Macapagal D., Murphy E., Mounts W., Berger-Bachi B., Projan S. 2004; Microarray-based analysis of the Staphylococcus aureus σB regulon. J Bacteriol 186:4085–4099 [View Article][PubMed]
    [Google Scholar]
  5. Boyle-Vavra S., Li X., Alam M. T., Read T. D., Sieth J., Cywes-Bentley C., Dobbins G., David M. Z., Kumar N., other authors. 2015; USA300 and USA500 clonal lineages of Staphylococcus aureus do not produce a capsular polysaccharide due to conserved mutations in the cap5 locus. MBio 6:e02585-14[PubMed] [CrossRef]
    [Google Scholar]
  6. Bronner S., Monteil H., Prevost G. 2004; Regulation of virulence determinants in Staphylococcus aureus: complexity and applications. FEMS Microbiol Rev 28:183–200 [View Article][PubMed]
    [Google Scholar]
  7. Charpentier E., Anton A. I., Barry P., Alfonso B., Fang Y., Novick R. P. 2004; Novel cassette-based shuttle vector system for gram-positive bacteria. Appl Environ Microbiol 70:6076–6085 [View Article][PubMed]
    [Google Scholar]
  8. Chen Z., Luong T. T., Lee C. Y. 2007; The sbcDC locus mediates repression of type 5 capsule production as part of the SOS response in Staphylococcus aureus . J Bacteriol 189:7343–7350 [View Article][PubMed]
    [Google Scholar]
  9. Chen K., Mizianty M. J., Kurgan L. 2012; Prediction and analysis of nucleotide-binding residues using sequence and sequence-derived structural descriptors. Bioinformatics 28:331–341 [View Article][PubMed]
    [Google Scholar]
  10. 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 [View Article][PubMed]
    [Google Scholar]
  11. Chini V., Foka A., Dimitracopoulos G., Spiliopoulou I. 2007; Absolute and relative real-time PCR in the quantification of tst gene expression among methicillin-resistant Staphylococcus aureus: evaluation by two mathematical models. Lett Appl Microbiol 45:479–484 [View Article][PubMed]
    [Google Scholar]
  12. Cocchiaro J. L., Gomez M. I., Risley A., Solinga R., Sordelli D. O., Lee J. C. 2006; Molecular characterization of the capsule locus from non-typeable Staphylococcus aureus . Mol Microbiol 59:948–960 [View Article][PubMed]
    [Google Scholar]
  13. Cunnion K. M., Lee J. C., Frank M. M. 2001; Capsule production and growth phase influence binding of complement to Staphylococcus aureus . Infect Immun 69:6796–6803 [View Article][PubMed]
    [Google Scholar]
  14. Dassy B., Fournier J. M. 1996; Respiratory activity is essential for post-exponential-phase production of type 5 capsular polysaccharide by Staphylococcus aureus . Infect Immun 64:2408–2414[PubMed]
    [Google Scholar]
  15. Dassy B., Hogan T., Foster T. J., Fournier J. M. 1993; Involvement of the accessory gene regulator (agr) in expression of type 5 capsular polysaccharide by Staphylococcus aureus . J Gen Microbiol 139:1301–1306 [View Article][PubMed]
    [Google Scholar]
  16. David M. Z., Daum R. S. 2010; Community-associated methicillin-resistant Staphylococcus aureus: epidemiology and clinical consequences of an emerging epidemic. Clin Microbiol Rev 23:616–687 [View Article][PubMed]
    [Google Scholar]
  17. Ding Y., Liu X., Chen F., Di H., Xu B., Zhou L., Deng X., Wu M., Yang C. G., Lan L. 2014; Metabolic sensor governing bacterial virulence in Staphylococcus aureus . Proc Natl Acad Sci U S A 111:E4981–E4990 [View Article][PubMed]
    [Google Scholar]
  18. Fattom A., Schneerson R., Watson D. C., Karakawa W. W., Fitzgerald D., Pastan I., Li X., Shiloach J., Bryla D. A., Robbins J. B. 1993; Laboratory and clinical evaluation of conjugate vaccines composed of Staphylococcus aureus type 5 and type 8 capsular polysaccharides bound to Pseudomonas aeruginosa recombinant exoprotein A. Infect Immun 61:1023–1032[PubMed]
    [Google Scholar]
  19. Fattom A. I., Sarwar J., Ortiz A., Naso R. 1996; Staphylococcus aureus capsular polysaccharide (CP) vaccine and CP-specific antibodies protect mice against bacterial challenge. Infect Immun 64:1659–1665[PubMed]
    [Google Scholar]
  20. 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 [View Article][PubMed]
    [Google Scholar]
  21. Graumann P., Wendrich T. M., Weber M. H., Schroder K., Marahiel M. A. 1997; A family of cold shock proteins in Bacillus subtilis is essential for cellular growth and for efficient protein synthesis at optimal and low temperatures. Mol Microbiol 25:741–756 [View Article][PubMed]
    [Google Scholar]
  22. Gupta R. K., Alba J., Xiong Y. Q., Bayer A. S., Lee C. Y. 2013; MgrA activates expression of capsule genes, but not the α-toxin gene in experimental Staphylococcus aureus endocarditis. J Infect Dis 208:1841–1848 [View Article][PubMed]
    [Google Scholar]
  23. Jutras B. L., Chenail A. M., Rowland C. L., Carroll D., Miller M. C., Bykowski T., Stevenson B. 2013; Eubacterial SpoVG homologs constitute a new family of site-specific DNA-binding proteins. PLoS One 8:e66683 [View Article][PubMed]
    [Google Scholar]
  24. Kampen A. H., Tollersrud T., Lund A. 2005; Staphylococcus aureus capsular polysaccharide types 5 and 8 reduce killing by bovine neutrophils in vitro. Infect Immun 73:1578–1583 [View Article][PubMed]
    [Google Scholar]
  25. Karakawa W. W., Vann W. F. 1982; Capsular polysaccharides of Staphylococcus aureus . Semin Infect Dis 4:285–293[PubMed]
    [Google Scholar]
  26. Karakawa W. W., Sutton A., Schneerson R., Karpas A., Vann W. F. 1988; Capsular antibodies induce type-specific phagocytosis of capsulated Staphylococcus aureus by human polymorphonuclear leukocytes. Infect Immun 56:1090–1095[PubMed]
    [Google Scholar]
  27. Lattar S. M., Noto Llana M., Denoel P., Germain S., Buzzola F. R., Lee J. C., Sordelli D. O. 2014; Protein antigens increase the protective efficacy of a capsule-based vaccine against Staphylococcus aureus in a rat model of osteomyelitis. Infect Immun 82:83–91 [View Article][PubMed]
    [Google Scholar]
  28. Lee J. C., Park J. S., Shepherd S. E., Carey V., Fattom A. 1997; Protective efficacy of antibodies to the Staphylococcus aureus type 5 capsular polysaccharide in a modified model of endocarditis in rats. Infect Immun 65:4146–4151[PubMed]
    [Google Scholar]
  29. Lei M. G., Lee C. Y. 2015; RbsR activates capsule but represses the rbsUDK operon in Staphylococcus aureus . J Bacteriol 197:3666–3675 [View Article][PubMed]
    [Google Scholar]
  30. Lowy F. D. 1998; Staphylococcus aureus infections. N Engl J Med 339:520–532 [View Article][PubMed]
    [Google Scholar]
  31. Lowy F. D. 2011; How Staphylococcus aureus adapts to its host. N Engl J Med 364:1987–1990 [View Article][PubMed]
    [Google Scholar]
  32. Luong T. T., Lee C. Y. 2002; Overproduction of type 8 capsular polysaccharide augments Staphylococcus aureus virulence. Infect Immun 70:3389–3395 [View Article][PubMed]
    [Google Scholar]
  33. Luong T. T., Lee C. Y. 2006; The arl locus positively regulates Staphylococcus aureus type 5 capsule via an mgrA-dependent pathway. Microbiology 152:3123–3131 [View Article][PubMed]
    [Google Scholar]
  34. Luong T., Sau S., Gomez M., Lee J. C., Lee C. Y. 2002; Regulation of Staphylococcus aureus capsular polysaccharide expression by agr and sarA . Infect Immun 70:444–450 [View Article][PubMed]
    [Google Scholar]
  35. Luong T. T., Newell S. W., Lee C. Y. 2003; Mgr, a novel global regulator in Staphylococcus aureus . J Bacteriol 185:3703–3710 [View Article][PubMed]
    [Google Scholar]
  36. Luong T. T., Sau K., Roux C., Sau S., Dunman P. M., Lee C. Y. 2011; Staphylococcus aureus ClpC divergently regulates capsule via sae and codY in strain Newman but activates capsule via codY in strain UAMS-1 and in strain Newman with repaired saeS . J Bacteriol 193:686–694 [View Article][PubMed]
    [Google Scholar]
  37. Majerczyk C. D., Dunman P. M., Luong T. T., Lee C. Y., Sadykov M. R., Somerville G. A., Bodi K., Sonenshein A. L. 2010; Direct targets of CodY in Staphylococcus aureus . J Bacteriol 192:2861–2877 [View Article][PubMed]
    [Google Scholar]
  38. Meier S., Goerke C., Wolz C., Seidl K., Homerova D., Schulthess B., Kormanec J., Berger-Bachi B., Bischoff M. 2007; σB and the σB-dependent arlRS and yabJ-spoVG loci affect capsule formation in Staphylococcus aureus . Infect Immun 75:4562–4571 [View Article][PubMed]
    [Google Scholar]
  39. Montgomery C. P., Boyle-Vavra S., Adem P. V., Lee J. C., Husain A. N., Clasen J., Daum R. S. 2008; Comparison of virulence in community-associated methicillin-resistant Staphylococcus aureus pulsotypes USA300 and USA400 in a rat model of pneumonia. J Infect Dis 198:561–570 [View Article][PubMed]
    [Google Scholar]
  40. Nanra J. S., Buitrago S. M., Crawford S., Ng J., Fink P. S., Hawkins J., Scully I. L., McNeil L. K., Aste-Amézaga J. M., other authors. 2013; Capsular polysaccharides are an important immune evasion mechanism for Staphylococcus aureus . Hum Vaccin Immunother 9:480–487 [View Article][PubMed]
    [Google Scholar]
  41. Nemeth J., Lee J. C. 1995; Antibodies to capsular polysaccharides are not protective against experimental Staphylococcus aureus endocarditis. Infect Immun 63:375–380[PubMed]
    [Google Scholar]
  42. Nilsson I. M., Lee J. C., Bremell T., Ryden C., Tarkowski A. 1997; The role of staphylococcal polysaccharide microcapsule expression in septicemia and septic arthritis. Infect Immun 65:4216–4221[PubMed]
    [Google Scholar]
  43. Novick R. P., Geisinger E. 2008; Quorum sensing in staphylococci. Annu Rev Genet 42:541–564 [View Article][PubMed]
    [Google Scholar]
  44. Nygaard T. K., Pallister K. B., Dumont A. L., Dewald M., Watkins R. L., Pallister E. Q., Malone C., Griffith S., Horswill A. R., other authors. 2012; Alpha-toxin induces programmed cell death of human T cells, B cells, and monocytes during USA300 infection. PLoS One 7:e36532 [View Article][PubMed]
    [Google Scholar]
  45. O'Riordan K., Lee J. C. 2004; Staphylococcus aureus capsular polysaccharides. Clin Microbiol Rev 17:218–234 [View Article][PubMed]
    [Google Scholar]
  46. Ouyang S., Sau S., Lee C. Y. 1999; Promoter analysis of the cap8 operon, involved in type 8 capsular polysaccharide production in Staphylococcus aureus . J Bacteriol 181:2492–2500[PubMed]
    [Google Scholar]
  47. Pané-Farré J., Jonas B., Förstner K., Engelmann S., Hecker M. 2006; The σB regulon in Staphylococcus aureus and its regulation. Int J Med Microbiol 296:237–258 [View Article][PubMed]
    [Google Scholar]
  48. Pohl K., Francois P., Stenz L., Schlink F., Geiger T., Herbert S., Goerke C., Schrenzel J., Wolz C. 2009; CodY in Staphylococcus aureus: a regulatory link between metabolism and virulence gene expression. J Bacteriol 191:2953–2963 [View Article][PubMed]
    [Google Scholar]
  49. Poutrel B., Gilbert F. B., Lebrun M. 1995; Effects of culture conditions on production of type 5 capsular polysaccharide by human and bovine Staphylococcus aureus strains. Clin Diagn Lab Immunol 2:166–171[PubMed]
    [Google Scholar]
  50. Ratnayake-Lecamwasam M., Serror P., Wong K. W., Sonenshein A. L. 2001; Bacillus subtilis CodY represses early-stationary-phase genes by sensing GTP levels. Genes Dev 15:1093–1103 [View Article][PubMed]
    [Google Scholar]
  51. Rost B., Yachdav G., Liu J. 2004; The PredictProtein server. Nucleic Acids Res 32:W321–W326 [View Article][PubMed]
    [Google Scholar]
  52. Sahukhal G. S., Elasri M. O. 2014; Identification and characterization of an operon, msaABCR, that controls virulence and biofilm development in Staphylococcus aureus . BMC Microbiol 14:154 [View Article][PubMed]
    [Google Scholar]
  53. Sahukhal G. S., Batte J. L., Elasri M. O. 2015; msaABCR operon positively regulates biofilm development by repressing proteases and autolysis in Staphylococcus aureus . FEMS Microbiol Lett 362:1–10 [View Article][PubMed]
    [Google Scholar]
  54. Samanta D., Elasri M. O. 2014; The msaABCR operon regulates resistance in vancomycin-intermediate Staphylococcus aureus strains. Antimicrob Agents Chemother 58:6685–6695 [View Article][PubMed]
    [Google Scholar]
  55. Sambanthamoorthy K., Smeltzer M. S., Elasri M. O. 2006; Identification and characterization of msa (SA1233), a gene involved in expression of SarA and several virulence factors in Staphylococcus aureus . Microbiology 152:2559–2572 [View Article][PubMed]
    [Google Scholar]
  56. Sambanthamoorthy K., Schwartz A., Nagarajan V., Elasri M. O. 2008; The role of msa in Staphylococcus aureus biofilm formation. BMC Microbiol 8:221 [View Article][PubMed]
    [Google Scholar]
  57. Sambrook J., Fritsch E. F., Maniatis T. 1989; Concentration of DNA solution. In Molecular Cloning: a Laboratory Manual p C1, 2nd edn.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  58. Sau S., Bhasin N., Wann E. R., Lee J. C., Foster T. J., Lee C. Y. 1997a; The Staphylococcus aureus allelic genetic loci for serotype 5 and 8 capsule expression contain the type-specific genes flanked by common genes. Microbiology 143:2395–2405 [View Article][PubMed]
    [Google Scholar]
  59. Sau S., Sun J., Lee C. Y. 1997b; Molecular characterization and transcriptional analysis of type 8 capsule genes in Staphylococcus aureus . J Bacteriol 179:1614–1621[PubMed]
    [Google Scholar]
  60. Schneider C. A., Rasband W. S., Eliceiri K. W. 2012; NIH Image to ImageJ: 25 years of image analysis. Nat Methods 9:671–675 [View Article][PubMed]
    [Google Scholar]
  61. Sengupta M., Jain V., Wilkinson B. J., Jayaswal R. K. 2012; Chromatin immunoprecipitation identifies genes under direct VraSR regulation in Staphylococcus aureus . Can J Microbiol 58:703–708 [View Article][PubMed]
    [Google Scholar]
  62. Shivers R. P., Sonenshein A. L. 2004; Activation of the Bacillus subtilis global regulator CodY by direct interaction with branched-chain amino acids. Mol Microbiol 53:599–611 [View Article][PubMed]
    [Google Scholar]
  63. Somerville G. A., Proctor R. A. 2009; At the crossroads of bacterial metabolism and virulence factor synthesis in staphylococci. Microbiol Mol Biol Rev 73:233–248 [View Article][PubMed]
    [Google Scholar]
  64. Sompolinsky D., Samra Z., Karakawa W. W., Vann W. F., Schneerson R., Malik Z. 1985; Encapsulation and capsular types in isolates of Staphylococcus aureus from different sources and relationship to phage types. J Clin Microbiol 22:828–834[PubMed]
    [Google Scholar]
  65. Sun F., Ji Q., Jones M. B., Deng X., Liang H., Frank B., Telser J., Peterson S. N., Bae T., He C. 2012; AirSR, a [2Fe-2S] cluster-containing two-component system, mediates global oxygen sensing and redox signaling in Staphylococcus aureus . J Am Chem Soc 134:305–314 [View Article][PubMed]
    [Google Scholar]
  66. Thakker M., Park J. S., Carey V., Lee J. C. 1998; Staphylococcus aureus serotype 5 capsular polysaccharide is antiphagocytic and enhances bacterial virulence in a murine bacteremia model. Infect Immun 66:5183–5189[PubMed]
    [Google Scholar]
  67. Tuchscherr L., Loffler B., Buzzola F. R., Sordelli D. O. 2010; Staphylococcus aureus adaptation to the host and persistence: role of loss of capsular polysaccharide expression. Future Microbiol 5:1823–1832 [View Article][PubMed]
    [Google Scholar]
  68. Voyich J. M., Braughton K. R., Sturdevant D. E., Whitney A. R., Said-Salim B., Porcella S. F., Long R. D., Dorward D. W., Gardner D. J., other authors. 2005; Insights into mechanisms used by Staphylococcus aureus to avoid destruction by human neutrophils. J Immunol 175:3907–3919 [View Article][PubMed]
    [Google Scholar]
  69. Wann E. R., Dassy B., Fournier J. M., Foster T. J. 1999; Genetic analysis of the cap5 locus of Staphylococcus aureus . FEMS Microbiol Lett 170:97–103 [View Article][PubMed]
    [Google Scholar]
  70. Watts A., Ke D., Wang Q., Pillay A., Nicholson-Weller A., Lee J. C. 2005; Staphylococcus aureus strains that express serotype 5 or serotype 8 capsular polysaccharides differ in virulence. Infect Immun 73:3502–3511 [View Article][PubMed]
    [Google Scholar]
  71. Zhao L., Xue T., Shang F., Sun H., Sun B. 2010; Staphylococcus aureus AI-2 quorum sensing associates with the KdpDE two-component system to regulate capsular polysaccharide synthesis and virulence. Infect Immun 78:3506–3515 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.000243
Loading
/content/journal/micro/10.1099/mic.0.000243
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