Skip to main content

Advertisement

Log in

Quorum sensing and alternative sigma factor RpoN regulate type VI secretion system I (T6SSVA1) in fish pathogen Vibrio alginolyticus

  • Short Communication
  • Published:
Archives of Microbiology Aims and scope Submit manuscript

Abstract

Type VI secretion system (T6SS) is a highly conserved bacterial protein secretion system and is precisely regulated in Gram-negative pathogens. In Vibrio alginolyticus, an important fish pathogen, two complete T6SS gene clusters (T6SSVA1 and T6SSVA2) were identified. In this study, expression of a hemolysin coregulated protein (Hcp1), which is one of the hallmarks of T6SS, was found to be strictly regulated in this bacterium. We showed that the expression of Hcp1 was growth phase-dependent and the production of Hcp1 reached a maximum in the exponential phase. The expression of Hcp1 was positively and negatively regulated by quorum sensing regulators LuxO and LuxR, respectively. In addition, we observed that Hcp1 expression required the alternative sigma factor RpoN and the enhancer-binding protein VasH, which is encoded in T6SSVA1 gene cluster. Moreover, LuxR, RpoN, and VasH could positively regulate the expression of other T6SS genes. Taken together, we demonstrated that the expression of T6SS in V. alginolyticus was under the regulation of quorum sensing and alternative sigma factor.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

References

  • Aubert DF, Flannagan RS, Valvano MA (2008) A novel sensor kinase-response regulator hybrid controls biofilm formation and Type VI secretion system activity in Burkholderia cenocepacia. Infe Immun 76:1979–1991

    Article  CAS  Google Scholar 

  • Bernard CS, Brunet YR, Gueguen E, Cascales E (2010) Nooks and crannies in type VI secretion regulation. J Bacteriol 192:3850–3860

    Article  PubMed  CAS  Google Scholar 

  • Bernard CS, Brunet YR, Gavioli M, Lloubès R, Cascales E (2011) Regulation of type VI secretion gene clusters by sigma54 and cognate enhancer binding proteins. J Bacteriol 193:2158–2167

    Article  PubMed  CAS  Google Scholar 

  • Bladergroen MR, Badelt K, Spaink HP (2003) Infection-blocking genes of a symbiotic Rhizobium leguminosarum strain that are involved in temperature-dependent protein secretion. Mol Plant-Microbe Interact 16:53–64

    Article  PubMed  CAS  Google Scholar 

  • Buck M, Gallegos MT, Studholme DJ, Guo Y, Gralla JD (2000) The bacterial enhancer-dependent sigma 54 (sigma N) transcription factor. J Bacteriol 182:4129–4136

    Article  PubMed  CAS  Google Scholar 

  • Chien JY, Shih JT, Hsueh PR, Yang PC, Luh KT (2002) Vibrio alginolyticus as the cause of pleural empyema and bacteremia in an immunocompromised patient. Eur J Clin Microbiol Infect Dis 21:401–403

    Article  PubMed  CAS  Google Scholar 

  • de Pace F, Boldrin de Paiva J, Nakazato G, Lancellotti M, Sircili MP, Guedes Stehling E et al (2011) Characterization of IcmF of the type VI secretion system in an avian pathogenic Escherichia coli (APEC) strain. Microbiology 157:2954–2962

    Article  PubMed  Google Scholar 

  • Denkin SM, Nelson DR (1999) Induction of protease activity in Vibrio anguillarum by gastrointestinal mucus. Appl Environ Microbiol 65:3555–3560

    PubMed  CAS  Google Scholar 

  • Dennis JJ, Zylstra GJ (1998) Plasposons: modular self-cloning minitransposon derivatives for rapid genetic analysis of gram-negative bacterial genomes. Appl Environ Microbiol 64:2710–2715

    PubMed  CAS  Google Scholar 

  • Ishikawa T, Rompikuntal PK, Lindmark B, Milton DL, Wai SN (2009) Quorum Sensing Regulation of the Two hcp Alleles in Vibrio cholerae O1 Strains. PLoS One 4:e6734

    Article  PubMed  Google Scholar 

  • Josenhans C, Suerbaum S (2002) The role of motility as a virulence factor in bacteria. Int J Med Microbiol 291:605–614

    Article  PubMed  CAS  Google Scholar 

  • Kawagishi I, Nakada M, Nishioka N, Homma M (1997) Cloning of a Vibrio alginolyticus rpoN gene that is required for polar flagellar formation. J Bacteriol 179:6851–6854

    PubMed  CAS  Google Scholar 

  • Kramer JA (2001) Omiga™: a PC-based sequence analysis tool. Mol Biotechnol 19:97–106

    Article  PubMed  CAS  Google Scholar 

  • Lee KK, Yu SR, Chen FR, Yang TI, Liu PC (1996a) Virulence of Vibrio alginolyticus isolated from diseased tiger prawn, Penaeus monodon. Curr Microbiol 32:229–231

    Article  PubMed  CAS  Google Scholar 

  • Lee KK, Yu SR, Yang TI, Liu PC, Chen FR (1996b) Isolation and characterization of Vibrio alginolyticus isolated from diseased kuruma prawn, Penaeus japonicus. Lett Appl Microbiol 22:111–114

    Article  PubMed  CAS  Google Scholar 

  • Lee KK, Yu SR, Liu PC (1997) Alkaline serine protease is an exotoxin of Vibrio alginolyticus in Kuruma prawn, Penaeus japonicus. Curr Microbiol 34:110–117

    Article  PubMed  Google Scholar 

  • Liang W, Wang S, Yu F, Zhang L, Qi G, Liu Y et al (2003) Construction and evaluation of a safe, live, oral Vibrio cholerae vaccine candidate, IEM108. Infect Immun 71:5498–5504

    Article  PubMed  CAS  Google Scholar 

  • Liu CH, Cheng W, Hsu JP, Chen JC (2004) Vibrio alginolyticus infection in the white shrimp Litopenaeus vannamei confirmed by polymerase chain reaction and 16S rDNA sequencing. Dis Aquat Org 61:169–174

    Article  PubMed  CAS  Google Scholar 

  • Liu H, Wang QY, Liu Q, Cao XD, Shi CB, Zhang YX (2011) Roles of Hfq in the stress adaptation and virulence in fish pathogen Vibrio alginolyticus and its potential application as a target for live attenuated vaccine. Appl Microbiol Biotechnol 91:353–364

    Article  PubMed  CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408

    Article  PubMed  CAS  Google Scholar 

  • Mattinen L, Somervuo P, Nykyri J, Nissinen R, Kouvonen P, Corthals G et al (2008) Microarray profiling of host-extract-induced genes and characterization of the type VI secretion cluster in the potato pathogen Pectobacterium atrosepticum. Microbiology 154:2387–2396

    Article  PubMed  CAS  Google Scholar 

  • Milton DL (2006) Quorum sensing in vibrios: complexity for diversification. Int J Med Microbiol 296:61–71

    Article  PubMed  CAS  Google Scholar 

  • Milton DL, Norqvist A, Wolf-Watz H (1992) Cloning of a metalloprotease gene involved in the virulence mechanism of Vibrio anguillarum. J Bacteriol 174:7235–7244

    PubMed  CAS  Google Scholar 

  • Morales VM, Bäckman A, Bagdasarian M (1991) A series of wide-host-range low-copy-number vectors that allow direct screening for recombinants. Gene 97:39–47

    Article  PubMed  CAS  Google Scholar 

  • Mougous JD, Cuff ME, Raunser S, Shen A, Zhou M, Gifford CA et al (2006) A virulence locus of Pseudomonas aeruginosa encodes a protein secretion apparatus. Science 312:1526–1530

    Article  PubMed  CAS  Google Scholar 

  • Mougous JD, Gifford CA, Ramsdell TL, Mekalanos JJ (2007) Threonine phosphorylation post-translationally regulates protein secretion in Pseudomonas aeruginosa. Nat Cell Biol 9:797–803

    Article  PubMed  CAS  Google Scholar 

  • Nicholas KB, Nicholas HB (1997) GeneDoc: a tool for editing and annotating multiple sequence alignments. http://www.psc.edu/biomed/genedoc

  • Parsons AD, Heffron F (2005) sciS, an icmF homolog in Salmonella enterica serovar typhimurium, limits intracellular replication and decreases virulence. Infect Immun 73:4338–4345

    Article  PubMed  CAS  Google Scholar 

  • Pieper R, Huang ST, Robinson JM, Clark DJ, Alami H, Parmar PP et al (2009) Temperature and growth phase influence the outer-membrane proteome and the expression of a type VI secretion system in Yersinia pestis. Microbiology 155:498–512

    Article  PubMed  CAS  Google Scholar 

  • Pukatzki S, Ma AT, Sturtevant D, Krastins B, Sarracino D, Nelson WC et al (2006) Identification of a conserved bacterial protein secretion system in Vibrio cholerae using the Dictyostelium host model system. Proc Natl Acad Sci USA 103:1528–1533

    Article  PubMed  CAS  Google Scholar 

  • Pukatzki S, Ma AT, Revel AT, Sturtevant D, Mekalanos JJ (2007) Type VI secretion system translocates a phage tail spike-like protein into target cells where it cross-links actin. Proc Natl Acad Sci USA 104:11508–11513

    Article  Google Scholar 

  • Rappas M, Bose D, Zhang XD (2006) Bacterial enhancer-binding proteins: unlocking σ54-dependent gene transcription. Curr Opin Struc Biol 17:110–116

    Article  Google Scholar 

  • Rui HP, Liu Q, Ma Y, Wang QY, Zhang YX (2008) Roles of LuxR in regulating extracellular alkaline serine protease A, extracellular polysaccharide and mobility of Vibrio alginolyticus. FEMS Microbiol Lett 285:155–162

    Article  PubMed  CAS  Google Scholar 

  • Rui HP, Liu Q, Wang QY, Ma Y, Liu H, Shi CB et al (2009) Role of alkaline serine protease, Asp, in Vibrio alginolyticus virulence and regulation of its expression by LuxO-LuxR regulatory system. J Microbiol Biotechnol 19:431–438

    Article  PubMed  CAS  Google Scholar 

  • Sambrook J, Fritsch E, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor

    Google Scholar 

  • Schell MA, Ulrich RL, Ribot WJ, Brueggemann EE, Hines HB, Chen D et al (2007) Type VI secretion is a major virulence determinant in Burkholderia mallei. Mol Microbiol 64:1466–1485

    Article  PubMed  CAS  Google Scholar 

  • Schumacher J, Joly N, Rappas M, Zhang XD, Buck M (2006) Structures and organization of AAA + enhancer binding proteins in transcriptional activation. J Struct Biol 156:190–199

    Article  PubMed  CAS  Google Scholar 

  • Spark RP, Fried ML, Perry C, Watkins C (1979) Vibrio alginolyticus wound infection: case report and review. Ann Clin Lab Sci 9:133–138

    PubMed  CAS  Google Scholar 

  • Srinivasa Rao PS, Yamada Y, Tan YP, Leung KY (2004) Use of proteomics to identify novel virulence determinants that are required for Edwardsiella tarda pathogenesis. Mol Microbiol 53:573–586

    Article  Google Scholar 

  • Suarez G, Sierra JC, Sha J, Wang S, Erova TE, Fadl AA et al (2008) Molecular characterization of a functional type VI secretion system from a clinical isolate of Aeromonas hydrophila. Microb Pathog 44:344–361

    Article  PubMed  CAS  Google Scholar 

  • Tian Y, Wang QY, Liu Q, Ma Y, Cao XD, Guan LY et al (2008) Involvement of LuxS in the regulation of motility and flagella biogenesis in Vibrio alginolyticus. Biosci Biotechnol Biochem 72:1063–1071

    Article  PubMed  CAS  Google Scholar 

  • Wang SY, Lauritz J, Jass J, Milton DL (2002) A ToxR homolog from Vibrio anguillarum serotype O1 regulates its own production, bile resistance, and biofilm formation. J Bacteriol 184:1630–1639

    Article  PubMed  CAS  Google Scholar 

  • Wang QY, Liu Q, Ma Y, Rui HP, Zhang YX (2007) LuxO controls extracellular protease, haemolytic activities and siderophore production in fish pathogen Vibrio alginolyticus. J Appl Microbiol 103:1525–1534

    Article  PubMed  CAS  Google Scholar 

  • Wang X, Wang QY, Xiao JF, Liu Q, Wu HZ, Xu LL et al (2009) Edwardsiella tarda T6SS component evpP is regulated by esrB and iron, and plays essential roles in the invasion of fish. Fish Shellfish Immunol 27:469–477

    Article  PubMed  CAS  Google Scholar 

  • Waters CM, Bassler BL (2005) Quorum sensing: cell-to-cell communication in bacteria. Annu Rev Cell Dev Biol 21:319–346

    Article  PubMed  CAS  Google Scholar 

  • Weber B, Hasic M, Chen C, Wai SN, Milton DL (2009) Type VI secretion modulates quorum sensing and stress response in Vibrio anguillarum. Environ Microbiol 11:3018–3028

    Article  PubMed  CAS  Google Scholar 

  • Wigneshweraraj S, Bose D, Burrows PC, Joly N, Schumacher J, Rappas M et al (2008) Modus operandi of the bacterial RNA polymerase containing the σ54 promoter-specificity factor. Mol Microbiol 68:538–546

    Article  PubMed  CAS  Google Scholar 

  • Williams SG, Varcoe LT, Attridge SR, Manning PA (1996) Vibrio cholerae Hcp, a secreted protein coregulated with HlyA. Infect Immun 64:283–289

    PubMed  CAS  Google Scholar 

  • Ye J, Ma Y, Liu Q, Zhao DL, Wang QY, Zhang YX (2008) Regulation of Vibrio alginolyticus virulence by the LuxS quorum-sensing system. J Fish Dis 31:161–169

    Article  PubMed  CAS  Google Scholar 

  • Yuan ZC, Liu P, Saenkham P, Kerr K, Nester EW (2008) Transcriptome profiling and functional analysis of Agrobacterium tumefaciens reveals a general conserved response to acidic conditions (pH 5.5) and a complex acid-mediated signaling involved in Agrobacterium-plant interactions. J Bacteriol 190:494–507

    Article  PubMed  CAS  Google Scholar 

  • Zhang W, Xu S, Li J, Shen X, Wang Y, Yuan Z (2011) Modulation of a thermoregulated type VI secretion system by AHL-dependent quorum sensing in Yersinia pseudotuberculosis. Arch Microbiol 193:351–363

    PubMed  CAS  Google Scholar 

  • Zhu J, Miller MB, Vance RE, Dziejman M, Bassler BL, Mekalanos JJ (2002) Quorum-sensing regulators control virulence gene expression in Vibrio cholerae. Proc Natl Acad Sci USA 99:3129–3134

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This study was supported by National Natural Science Foundation of China (No. 30901113) and Shanghai Leading Academic Discipline Project (No. B505).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Qiyao Wang.

Additional information

Communicated by Shuang-Jiang Liu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sheng, L., Gu, D., Wang, Q. et al. Quorum sensing and alternative sigma factor RpoN regulate type VI secretion system I (T6SSVA1) in fish pathogen Vibrio alginolyticus . Arch Microbiol 194, 379–390 (2012). https://doi.org/10.1007/s00203-011-0780-z

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00203-011-0780-z

Keywords

Navigation