1887

Abstract

A single polar flagellum and motility are potential virulence factors of , a foodborne pathogen. In the present study, the functions of FlhF and regulatory characteristics of the expression of were investigated. A deletion mutation in abolished motility, flagella formation and flagellin synthesis, and introduction of complemented the defects. The mutant revealed decreased expression of the class III and IV flagella genes, indicating that FlhF is a key regulator for the flagellar biogenesis of . The influence of global regulatory proteins on the expression of was examined and SmcR, a LuxR homologue, was found to downregulate expression at the transcriptional level. SmcR represses expression only in the stationary phase of growth and exerts its effects by directly binding to the promoter region. Finally, an SmcR binding site, centred at 22.5 bp upstream of the transcription start site, was identified by a DNase I protection assay. The combined results demonstrate that a quorum sensing master regulator SmcR influences the motility and flagellar biogenesis of through modulating the expression of FlhF in a growth-phase-dependent manner.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.059071-0
2012-08-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/158/8/2017.html?itemId=/content/journal/micro/10.1099/mic.0.059071-0&mimeType=html&fmt=ahah

References

  1. Beyhan S., Bilecen K., Salama S. R., Casper-Lindley C., Yildiz F. H. ( 2007). Regulation of rugosity and biofilm formation in Vibrio cholerae: comparison of VpsT and VpsR regulons and epistasis analysis of vpsT, vpsR, and hapR . J Bacteriol 189:388–402 [View Article][PubMed]
    [Google Scholar]
  2. Connell T. D., Martone A. J., Holmes R. K. ( 1995). A new mobilizable cosmid vector for use in Vibrio cholerae and other gram− bacteria. Gene 153:85–87 [View Article][PubMed]
    [Google Scholar]
  3. Correa N. E., Peng F., Klose K. E. ( 2005). Roles of the regulatory proteins FlhF and FlhG in the Vibrio cholerae flagellar transcription hierarchy. J Bacteriol 187:6324–6332 [View Article][PubMed]
    [Google Scholar]
  4. Croxatto A., Chalker V. J., Lauritz J., Jass J., Hardman A., Williams P., Cámara M., Milton D. L. ( 2002). VanT, a homologue of Vibrio harveyi LuxR, regulates serine, metalloprotease, pigment, and biofilm production in Vibrio anguillarum . J Bacteriol 184:1617–1629 [View Article][PubMed]
    [Google Scholar]
  5. Fuqua C., Greenberg E. P. ( 2002). Listening in on bacteria: acyl-homoserine lactone signalling. Nat Rev Mol Cell Biol 3:685–695 [View Article][PubMed]
    [Google Scholar]
  6. Ghelardi E., Celandroni F., Salvetti S., Beecher D. J., Gominet M., Lereclus D., Wong A. C., Senesi S. ( 2002). Requirement of flhA for swarming differentiation, flagellin export, and secretion of virulence-associated proteins in Bacillus thuringiensis . J Bacteriol 184:6424–6433 [View Article][PubMed]
    [Google Scholar]
  7. Green J. C., Kahramanoglou C., Rahman A., Pender A. M., Charbonnel N., Fraser G. M. ( 2009). Recruitment of the earliest component of the bacterial flagellum to the old cell division pole by a membrane-associated signal recognition particle family GTP-binding protein. J Mol Biol 391:679–690 [View Article][PubMed]
    [Google Scholar]
  8. Guzman L. M., Belin D., Carson M. J., Beckwith J. ( 1995). Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter. J Bacteriol 177:4121–4130[PubMed]
    [Google Scholar]
  9. Hayashi F., Smith K. D., Ozinsky A., Hawn T. R., Yi E. C., Goodlett D. R., Eng J. K., Akira S., Underhill D. M., Aderem A. ( 2001). The innate immune response to bacterial flagellin is mediated by Toll-like receptor 5. Nature 410:1099–1103 [View Article][PubMed]
    [Google Scholar]
  10. Jeong H. S., Jeong K. C., Choi H. K., Park K. J., Lee K. H., Rhee J. H., Choi S. H. ( 2001). Differential expression of Vibrio vulnificus elastase gene in a growth phase-dependent manner by two different types of promoters. J Biol Chem 276:13875–13880[PubMed]
    [Google Scholar]
  11. Jeong H. S., Lee M. H., Lee K. H., Park S. J., Choi S. H. ( 2003a). SmcR and cyclic AMP receptor protein coactivate Vibrio vulnificus vvpE encoding elastase through the RpoS-dependent promoter in a synergistic manner. J Biol Chem 278:45072–45081 [View Article][PubMed]
    [Google Scholar]
  12. Jeong H. S., Rhee J. E., Lee J. H., Choi H. K., Kim D. I., Lee M. H., Park S. J., Choi S. H. ( 2003b). Identification of Vibrio vulnificus lrp and its influence on survival under various stresses. J Microbiol Biotechnol 13:159–163
    [Google Scholar]
  13. Jeong H. S., Kim S. M., Lim M. S., Kim K. S., Choi S. H. ( 2010). Direct interaction between quorum-sensing regulator SmcR and RNA polymerase is mediated by integration host factor to activate vvpE encoding elastase in Vibrio vulnificus . J Biol Chem 285:9357–9366 [View Article][PubMed]
    [Google Scholar]
  14. Jobling M. G., Holmes R. K. ( 1997). Characterization of hapR, a positive regulator of the Vibrio cholerae HA/protease gene hap, and its identification as a functional homologue of the Vibrio harveyi luxR gene. Mol Microbiol 26:1023–1034 [View Article][PubMed]
    [Google Scholar]
  15. Jones M. K., Oliver J. D. ( 2009). Vibrio vulnificus: disease and pathogenesis. Infect Immun 77:1723–1733 [View Article][PubMed]
    [Google Scholar]
  16. Kim Y. R., Rhee J. H. ( 2003). Flagellar basal body flg operon as a virulence determinant of Vibrio vulnificus . Biochem Biophys Res Commun 304:405–410 [View Article][PubMed]
    [Google Scholar]
  17. Klose K. E., Mekalanos J. J. ( 1998). Differential regulation of multiple flagellins in Vibrio cholerae . J Bacteriol 180:303–316[PubMed]
    [Google Scholar]
  18. Kojima M., Nishioka N., Kusumoto A., Yagasaki J., Fukuda T., Homma M. ( 2011). Conversion of mono-polar to peritrichous flagellation in Vibrio alginolyticus . Microbiol Immunol 55:76–83 [View Article][PubMed]
    [Google Scholar]
  19. Kusumoto A., Kamisaka K., Yakushi T., Terashima H., Shinohara A., Homma M. ( 2006). Regulation of polar flagellar number by the flhF and flhG genes in Vibrio alginolyticus . J Biochem 139:113–121 [View Article][PubMed]
    [Google Scholar]
  20. Kusumoto A., Shinohara A., Terashima H., Kojima S., Yakushi T., Homma M. ( 2008). Collaboration of FlhF and FlhG to regulate polar-flagella number and localization in Vibrio alginolyticus . Microbiology 154:1390–1399 [View Article][PubMed]
    [Google Scholar]
  21. Kusumoto A., Nishioka N., Kojima S., Homma M. ( 2009). Mutational analysis of the GTP-binding motif of FlhF which regulates the number and placement of the polar flagellum in Vibrio alginolyticus . J Biochem 146:643–650 [View Article][PubMed]
    [Google Scholar]
  22. Lee J. H., Rho J. B., Park K. J., Kim C. B., Han Y. S., Choi S. H., Lee K. H., Park S. J. ( 2004). Role of flagellum and motility in pathogenesis of Vibrio vulnificus . Infect Immun 72:4905–4910 [View Article][PubMed]
    [Google Scholar]
  23. Lee S. E., Kim S. Y., Jeong B. C., Kim Y. R., Bae S. J., Ahn O. S., Lee J. J., Song H. C., Kim J. M. & other authors ( 2006). A bacterial flagellin, Vibrio vulnificus FlaB, has a strong mucosal adjuvant activity to induce protective immunity. Infect Immun 74:694–702 [View Article][PubMed]
    [Google Scholar]
  24. Lee D. H., Jeong H. S., Jeong H. G., Kim K. M., Kim H., Choi S. H. ( 2008). A consensus sequence for binding of SmcR, a Vibrio vulnificus LuxR homologue, and genome-wide identification of the SmcR regulon. J Biol Chem 283:23610–23618 [View Article][PubMed]
    [Google Scholar]
  25. Liu Z., Miyashiro T., Tsou A., Hsiao A., Goulian M., Zhu J. ( 2008). Mucosal penetration primes Vibrio cholerae for host colonization by repressing quorum sensing. Proc Natl Acad Sci U S A 105:9769–9774 [View Article][PubMed]
    [Google Scholar]
  26. McCarter L. L. ( 1998). OpaR, a homolog of Vibrio harveyi LuxR, controls opacity of Vibrio parahaemolyticus . J Bacteriol 180:3166–3173[PubMed]
    [Google Scholar]
  27. McCarter L. L. ( 2001). Polar flagellar motility of the Vibrionaceae . Microbiol Mol Biol Rev 65:445–462 [View Article][PubMed]
    [Google Scholar]
  28. McDougald D., Rice S. A., Kjelleberg S. ( 2001). SmcR-dependent regulation of adaptive phenotypes in Vibrio vulnificus . J Bacteriol 183:758–762 [View Article][PubMed]
    [Google Scholar]
  29. Miller V. L., Mekalanos J. J. ( 1988). A novel suicide vector and its use in construction of insertion mutations: osmoregulation of outer membrane proteins and virulence determinants in Vibrio cholerae requires toxR . J Bacteriol 170:2575–2583[PubMed]
    [Google Scholar]
  30. Moisi M., Jenul C., Butler S. M., New A., Tutz S., Reidl J., Klose K. E., Camilli A., Schild S. ( 2009). A novel regulatory protein involved in motility of Vibrio cholerae . J Bacteriol 191:7027–7038 [View Article][PubMed]
    [Google Scholar]
  31. Nakhamchik A., Wilde C., Rowe-Magnus D. A. ( 2008). Cyclic-di-GMP regulates extracellular polysaccharide production, biofilm formation, and rugose colony development by Vibrio vulnificus . Appl Environ Microbiol 74:4199–4209 [View Article][PubMed]
    [Google Scholar]
  32. Ng W. L., Bassler B. L. ( 2009). Bacterial quorum-sensing network architectures. Annu Rev Genet 43:197–222 [View Article][PubMed]
    [Google Scholar]
  33. Philippe N., Alcaraz J. P., Coursange E., Geiselmann J., Schneider D. ( 2004). Improvement of pCVD442, a suicide plasmid for gene allele exchange in bacteria. Plasmid 51:246–255 [View Article][PubMed]
    [Google Scholar]
  34. Kim Y. R., Rhee J. H. ( 2003). Flagellar basal body flg operon as a virulence determinant of Vibrio vulnificus . Biochem Biophys Res Commun 304:405–410 [View Article][PubMed]
    [Google Scholar]
  35. Sambrook J., Russell D. W. 2001 Molecular Cloning: a Laboratory Manual, 3rd edn.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  36. Smith K. D., Andersen-Nissen E., Hayashi F., Strobe K., Bergman M. A., Barrett S. L., Cookson B. T., Aderem A. ( 2003). Toll-like receptor 5 recognizes a conserved site on flagellin required for protofilament formation and bacterial motility. Nat Immunol 4:1247–1253 [View Article][PubMed]
    [Google Scholar]
  37. Sultan S. Z., Silva A. J., Benitez J. A. ( 2010). The PhoB regulatory system modulates biofilm formation and stress response in El Tor biotype Vibrio cholerae . FEMS Microbiol Lett 302:22–31 [View Article][PubMed]
    [Google Scholar]
  38. Tian Y., Wang Q., Liu Q., Ma Y., Cao X., Guan L., Zhang Y. ( 2008). Involvement of LuxS in the regulation of motility and flagella biogenesis in Vibrio alginolyticus . Biosci Biotechnol Biochem 72:1063–1071 [View Article][PubMed]
    [Google Scholar]
  39. Waters C. M., Bassler B. L. ( 2006). The Vibrio harveyi quorum-sensing system uses shared regulatory components to discriminate between multiple autoinducers. Genes Dev 20:2754–2767 [View Article][PubMed]
    [Google Scholar]
  40. Wright A. C., Simpson L. M., Oliver J. D., Morris J. G. Jr ( 1990). Phenotypic evaluation of acapsular transposon mutants of Vibrio vulnificus . Infect Immun 58:1769–1773[PubMed]
    [Google Scholar]
  41. Zhu J., Miller M. B., Vance R. E., Dziejman M., Bassler B. L., Mekalanos J. J. ( 2002). Quorum-sensing regulators control virulence gene expression in Vibrio cholerae . Proc Natl Acad Sci U S A 99:3129–3134 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.059071-0
Loading
/content/journal/micro/10.1099/mic.0.059071-0
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