#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

Response Regulator VxrB Controls Colonization and Regulates the Type VI Secretion System


Pathogenic bacteria experience varying conditions during infection of human hosts and often use two-component signal transduction systems (TCSs) to monitor their environment. TCS consists of a histidine kinase (HK), which senses environmental signals, and a corresponding response regulator (RR), which mediates a cellular response. The genome of the human pathogen V. cholerae contains a multitude of genes encoding HKs and RRs proteins. In the present study, we systematically analyzed the role of each V. cholerae RR for its role in pathogenesis. We identified a previously uncharacterized RR, VxrB, as a new virulence factor. We demonstrated that VxrB controls expression of the type VI secretion system (T6SS), a virulence nanomachine that directly translocates effectors into bacterial or host cells, thereby facilitating colonization by competing with sister cells and intestinal microbiota. This study represents the first systematic analysis of the role of all RRs in V. cholerae pathogenesis and provides a foundation for understanding the signal transduction pathways controlling V. cholerae pathogenesis.


Vyšlo v časopise: Response Regulator VxrB Controls Colonization and Regulates the Type VI Secretion System. PLoS Pathog 11(5): e32767. doi:10.1371/journal.ppat.1004933
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1004933

Souhrn

Pathogenic bacteria experience varying conditions during infection of human hosts and often use two-component signal transduction systems (TCSs) to monitor their environment. TCS consists of a histidine kinase (HK), which senses environmental signals, and a corresponding response regulator (RR), which mediates a cellular response. The genome of the human pathogen V. cholerae contains a multitude of genes encoding HKs and RRs proteins. In the present study, we systematically analyzed the role of each V. cholerae RR for its role in pathogenesis. We identified a previously uncharacterized RR, VxrB, as a new virulence factor. We demonstrated that VxrB controls expression of the type VI secretion system (T6SS), a virulence nanomachine that directly translocates effectors into bacterial or host cells, thereby facilitating colonization by competing with sister cells and intestinal microbiota. This study represents the first systematic analysis of the role of all RRs in V. cholerae pathogenesis and provides a foundation for understanding the signal transduction pathways controlling V. cholerae pathogenesis.


Zdroje

1. Ali M, Lopez AL, You YA, Kim YE, Sah B, Maskery B, et al. The global burden of cholera. Bulletin of the World Health Organization. 2012 Mar 1;90(3):209–18A. doi: 10.2471/BLT.11.093427 22461716

2. Kaper JB, Morris JG Jr., Levine MM. Cholera. Clin Microbiol Rev. 1995 Jan;8(1):48–86. 7704895

3. DiRita VJ, Parsot C, Jander G, Mekalanos JJ. Regulatory cascade controls virulence in Vibrio cholerae. Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5403–7. 2052618

4. Higgins DE, Nazareno E, DiRita VJ. The virulence gene activator ToxT from Vibrio cholerae is a member of the AraC family of transcriptional activators. J Bacteriol. 1992 Nov;174(21):6974–80. 1400247

5. Higgins DE, DiRita VJ. Transcriptional control of toxT, a regulatory gene in the ToxR regulon of Vibrio cholerae. Mol Microbiol. 1994 Oct;14(1):17–29. 7830555

6. Hase CC, Mekalanos JJ. TcpP protein is a positive regulator of virulence gene expression in Vibrio cholerae. Proc Natl Acad Sci U S A. 1998 Jan 20;95(2):730–4. 9435261

7. Krukonis ES, Yu RR, Dirita VJ. The Vibrio cholerae ToxR/TcpP/ToxT virulence cascade: distinct roles for two membrane-localized transcriptional activators on a single promoter. Mol Microbiol. 2000 Oct;38(1):67–84. 11029691

8. Kovacikova G, Lin W, Skorupski K. Vibrio cholerae AphA uses a novel mechanism for virulence gene activation that involves interaction with the LysR-type regulator AphB at the tcpPH promoter. Mol Microbiol. 2004 Jul;53(1):129–42. 15225309

9. Skorupski K, Taylor RK. A new level in the Vibrio cholerae ToxR virulence cascade: AphA is required for transcriptional activation of the tcpPH operon. Mol Microbiol. 1999 Feb;31(3):763–71. 10048021

10. Kovacikova G, Skorupski K. Regulation of virulence gene expression in Vibrio cholerae by quorum sensing: HapR functions at the aphA promoter. Mol Microbiol. 2002 Nov;46(4):1135–47. 12421317

11. Pukatzki S, Ma AT, Sturtevant D, Krastins B, Sarracino D, Nelson WC, et al. Identification of a conserved bacterial protein secretion system in Vibrio cholerae using the Dictyostelium host model system. Proc Natl Acad Sci U S A. 2006 Jan 31;103(5):1528–33. 16432199

12. Ho BT, Dong TG, Mekalanos JJ. A view to a kill: the bacterial type VI secretion system. Cell host & microbe. 2014 Jan 15;15(1):9–21.

13. Silverman JM, Brunet YR, Cascales E, Mougous JD. Structure and regulation of the type VI secretion system. Annu Rev Microbiol. 2012;66:453–72. doi: 10.1146/annurev-micro-121809-151619 22746332

14. Pukatzki S, Ma AT, Revel AT, Sturtevant D, Mekalanos JJ. Type VI secretion system translocates a phage tail spike-like protein into target cells where it cross-links actin. Proc Natl Acad Sci U S A. 2007 Sep 25;104(39):15508–13. 17873062

15. Dong TG, Ho BT, Yoder-Himes DR, Mekalanos JJ. Identification of T6SS-dependent effector and immunity proteins by Tn-seq in Vibrio cholerae. Proc Natl Acad Sci U S A. 2013 Feb 12;110(7):2623–8. doi: 10.1073/pnas.1222783110 23362380

16. Brooks TM, Unterweger D, Bachmann V, Kostiuk B, Pukatzki S. Lytic activity of the Vibrio cholerae type VI secretion toxin VgrG-3 is inhibited by the antitoxin TsaB. J Biol Chem. 2013 Mar 15;288(11):7618–25. doi: 10.1074/jbc.M112.436725 23341465

17. Ma AT, McAuley S, Pukatzki S, Mekalanos JJ. Translocation of a Vibrio cholerae type VI secretion effector requires bacterial endocytosis by host cells. Cell host & microbe. 2009 Mar 19;5(3):234–43.

18. Russell AB, LeRoux M, Hathazi K, Agnello DM, Ishikawa T, Wiggins PA, et al. Diverse type VI secretion phospholipases are functionally plastic antibacterial effectors. Nature. 2013 Apr 25;496(7446):508–12. doi: 10.1038/nature12074 23552891

19. Miyata ST, Kitaoka M, Brooks TM, McAuley SB, Pukatzki S. Vibrio cholerae requires the type VI secretion system virulence factor VasX to kill Dictyostelium discoideum. Infect Immun. 2011 Jul;79(7):2941–9. doi: 10.1128/IAI.01266-10 21555399

20. Miyata ST, Unterweger D, Rudko SP, Pukatzki S. Dual expression profile of type VI secretion system immunity genes protects pandemic Vibrio cholerae. PLoS Pathog. 2013;9(12):e1003752. doi: 10.1371/journal.ppat.1003752 24348240

21. Unterweger D, Miyata ST, Bachmann V, Brooks TM, Mullins T, Kostiuk B, et al. The Vibrio cholerae type VI secretion system employs diverse effector modules for intraspecific competition. Nature communications. 2014;5:3549. doi: 10.1038/ncomms4549 24686479

22. Basler M, Pilhofer M, Henderson GP, Jensen GJ, Mekalanos JJ. Type VI secretion requires a dynamic contractile phage tail-like structure. Nature. 2012 Mar 8;483(7388):182–6. doi: 10.1038/nature10846 22367545

23. Zheng J, Ho B, Mekalanos JJ. Genetic analysis of anti-amoebae and anti-bacterial activities of the type VI secretion system in Vibrio cholerae. PloS one. 2011;6(8):e23876. doi: 10.1371/journal.pone.0023876 21909372

24. Kitaoka M, Miyata ST, Brooks TM, Unterweger D, Pukatzki S. VasH is a transcriptional regulator of the type VI secretion system functional in endemic and pandemic Vibrio cholerae. J Bacteriol. 2011 Dec;193(23):6471–82. doi: 10.1128/JB.05414-11 21949076

25. Bernard CS, Brunet YR, Gavioli M, Lloubes R, Cascales E. Regulation of type VI secretion gene clusters by sigma54 and cognate enhancer binding proteins. J Bacteriol. 2011 May;193(9):2158–67. doi: 10.1128/JB.00029-11 21378190

26. Dong TG, Mekalanos JJ. Characterization of the RpoN regulon reveals differential regulation of T6SS and new flagellar operons in Vibrio cholerae O37 strain V52. Nucleic Acids Res. 2012 Sep;40(16):7766–75. doi: 10.1093/nar/gks567 22723378

27. Zheng J, Shin OS, Cameron DE, Mekalanos JJ. Quorum sensing and a global regulator TsrA control expression of type VI secretion and virulence in Vibrio cholerae. Proc Natl Acad Sci U S A. 2010 Dec 7;107(49):21128–33. doi: 10.1073/pnas.1014998107 21084635

28. Ishikawa T, Rompikuntal PK, Lindmark B, Milton DL, Wai SN. Quorum sensing regulation of the two hcp alleles in Vibrio cholerae O1 strains. PloS one. 2009;4(8):e6734. doi: 10.1371/journal.pone.0006734 19701456

29. Ma AT, Mekalanos JJ. In vivo actin cross-linking induced by Vibrio cholerae type VI secretion system is associated with intestinal inflammation. Proc Natl Acad Sci U S A. 2010 Mar 2;107(9):4365–70. doi: 10.1073/pnas.0915156107 20150509

30. Fu Y, Waldor MK, Mekalanos JJ. Tn-Seq analysis of Vibrio cholerae intestinal colonization reveals a role for T6SS-mediated antibacterial activity in the host. Cell host & microbe. 2013 Dec 11;14(6):652–63.

31. Beier D, Gross R. Regulation of bacterial virulence by two-component systems. Curr Opin Microbiol. 2006 Apr;9(2):143–52. 16481212

32. Krell T, Lacal J, Busch A, Silva-Jimenez H, Guazzaroni ME, Ramos JL. Bacterial sensor kinases: diversity in the recognition of environmental signals. Annu Rev Microbiol. 2010;64:539–59. doi: 10.1146/annurev.micro.112408.134054 20825354

33. Galperin MY. Diversity of structure and function of response regulator output domains. Curr Opin Microbiol. 2010 Apr;13(2):150–9. doi: 10.1016/j.mib.2010.01.005 20226724

34. Gao R, Stock AM. Biological insights from structures of two-component proteins. Annu Rev Microbiol. 2009;63:133–54. doi: 10.1146/annurev.micro.091208.073214 19575571

35. Laub MT, Goulian M. Specificity in two-component signal transduction pathways. Annual review of genetics. 2007;41:121–45. 18076326

36. Jang J, Jung KT, Park J, Yoo CK, Rhie GE. The Vibrio cholerae VarS/VarA two-component system controls the expression of virulence proteins through ToxT regulation. Microbiology. 2011 May;157(Pt 5):1466–73. doi: 10.1099/mic.0.043737-0 21330435

37. Zhu J, Miller MB, Vance RE, Dziejman M, Bassler BL, Mekalanos JJ. Quorum-sensing regulators control virulence gene expression in Vibrio cholerae. Proc Natl Acad Sci U S A. 2002 Mar 5;99(5):3129–34. 11854465

38. Tischler AD, Camilli A. Cyclic diguanylate regulates Vibrio cholerae virulence gene expression. Infect Immun. 2005 Sep;73(9):5873–82. 16113306

39. Tischler AD, Lee SH, Camilli A. The Vibrio cholerae vieSAB locus encodes a pathway contributing to cholera toxin production. J Bacteriol. 2002 Aug;184(15):4104–13. 12107127

40. Pratt JT, Ismail AM, Camilli A. PhoB regulates both environmental and virulence gene expression in Vibrio cholerae. Mol Microbiol. 2010 Sep;77(6):1595–605. doi: 10.1111/j.1365-2958.2010.07310.x 20659293

41. Sengupta N, Paul K, Chowdhury R. The global regulator ArcA modulates expression of virulence factors in Vibrio cholerae. Infect Immun. 2003 Oct;71(10):5583–9. 14500477

42. Bilecen K, Fong JC, Cheng A, Jones CJ, Zamorano-Sanchez D, Yildiz FH. Polymyxin B Resistance and biofilm formation in Vibrio cholerae is controlled by the response regulator CarR. Infect Immun. 2015 Jan 12.

43. Butler SM, Camilli A. Both chemotaxis and net motility greatly influence the infectivity of Vibrio cholerae. Proc Natl Acad Sci U S A. 2004 Apr 6;101(14):5018–23. 15037750

44. Correa NE, Lauriano CM, McGee R, Klose KE. Phosphorylation of the flagellar regulatory protein FlrC is necessary for Vibrio cholerae motility and enhanced colonization. Mol Microbiol. 2000 Feb;35(4):743–55. 10692152

45. Butler SM, Camilli A. Going against the grain: chemotaxis and infection in Vibrio cholerae. Nature reviews Microbiology. 2005 Aug;3(8):611–20. 16012515

46. Taylor RK, Miller VL, Furlong DB, Mekalanos JJ. Use of phoA gene fusions to identify a pilus colonization factor coordinately regulated with cholera toxin. Proc Natl Acad Sci U S A. 1987 May;84(9):2833–7. 2883655

47. Galperin MY. Structural classification of bacterial response regulators: diversity of output domains and domain combinations. J Bacteriol. 2006 Jun;188(12):4169–82. 16740923

48. Delgado J, Forst S, Harlocker S, Inouye M. Identification of a phosphorylation site and functional analysis of conserved aspartic acid residues of OmpR, a transcriptional activator for ompF and ompC in Escherichia coli. Mol Microbiol. 1993 Dec;10(5):1037–47. 7934854

49. Ma S, Selvaraj U, Ohman DE, Quarless R, Hassett DJ, Wozniak DJ. Phosphorylation-independent activity of the response regulators AlgB and AlgR in promoting alginate biosynthesis in mucoid Pseudomonas aeruginosa. J Bacteriol. 1998 Feb;180(4):956–68. 9473053

50. Ishikawa T, Sabharwal D, Broms J, Milton DL, Sjostedt A, Uhlin BE, et al. Pathoadaptive conditional regulation of the type VI secretion system in Vibrio cholerae O1 strains. Infect Immun. 2012 Feb;80(2):575–84. doi: 10.1128/IAI.05510-11 22083711

51. Hussa EA, O'Shea TM, Darnell CL, Ruby EG, Visick KL. Two-component response regulators of Vibrio fischeri: identification, mutagenesis, and characterization. J Bacteriol. 2007 Aug;189(16):5825–38. 17586650

52. Kobayashi K, Ogura M, Yamaguchi H, Yoshida K, Ogasawara N, Tanaka T, et al. Comprehensive DNA microarray analysis of Bacillus subtilis two-component regulatory systems. J Bacteriol. 2001 Dec;183(24):7365–70. 11717295

53. Lange R, Wagner C, de Saizieu A, Flint N, Molnos J, Stieger M, et al. Domain organization and molecular characterization of 13 two-component systems identified by genome sequencing of Streptococcus pneumoniae. Gene. 1999 Sep 3;237(1):223–34. 10524254

54. Throup JP, Koretke KK, Bryant AP, Ingraham KA, Chalker AF, Ge Y, et al. A genomic analysis of two-component signal transduction in Streptococcus pneumoniae. Mol Microbiol. 2000 Feb;35(3):566–76. 10672179

55. Yamamoto K, Hirao K, Oshima T, Aiba H, Utsumi R, Ishihama A. Functional characterization in vitro of all two-component signal transduction systems from Escherichia coli. J Biol Chem. 2005 Jan 14;280(2):1448–56. 15522865

56. Mandlik A, Livny J, Robins WP, Ritchie JM, Mekalanos JJ, Waldor MK. RNA-Seq-based monitoring of infection-linked changes in Vibrio cholerae gene expression. Cell host & microbe. 2011 Aug 18;10(2):165–74.

57. Kamp HD, Patimalla-Dipali B, Lazinski DW, Wallace-Gadsden F, Camilli A. Gene fitness landscapes of Vibrio cholerae at important stages of its life cycle. PLoS Pathog. 2013;9(12):e1003800. doi: 10.1371/journal.ppat.1003800 24385900

58. Kelley LA, Sternberg MJ. Protein structure prediction on the Web: a case study using the Phyre server. Nature protocols. 2009;4(3):363–71. doi: 10.1038/nprot.2009.2 19247286

59. Bernard CS, Brunet YR, Gueguen E, Cascales E. Nooks and crannies in type VI secretion regulation. J Bacteriol. 2010 Aug;192(15):3850–60. doi: 10.1128/JB.00370-10 20511495

60. Borgeaud S, Metzger LC, Scrignari T, Blokesch M. Bacterial evolution. The type VI secretion system of Vibrio cholerae fosters horizontal gene transfer. Science. 2015 Jan 2;347(6217):63–7. doi: 10.1126/science.1260064 25554784

61. National Research Council (U.S.). Committee for the Update of the Guide for the Care and Use of Laboratory Animals., Institute for Laboratory Animal Research (U.S.), National Academies Press (U.S.). Guide for the care and use of laboratory animals. 8th ed. Washington, D.C.: National Academies Press; 2011. xxv, 220 p. p.

62. Lim B, Beyhan S, Meir J, Yildiz FH. Cyclic-diGMP signal transduction systems in Vibrio cholerae: modulation of rugosity and biofilm formation. Mol Microbiol. 2006 Apr;60(2):331–48. 16573684

63. Iwanaga M, Yamamoto K, Higa N, Ichinose Y, Nakasone N, Tanabe M. Culture conditions for stimulating cholera toxin production by Vibrio cholerae O1 El Tor. Microbiology and immunology. 1986;30(11):1075–83. 3543624

64. Fong JC, Yildiz FH. The rbmBCDEF gene cluster modulates development of rugose colony morphology and biofilm formation in Vibrio cholerae. J Bacteriol. 2007 Mar;189(6):2319–30. 17220218

65. Lefebvre B, Formstecher P, Lefebvre P. Improvement of the gene splicing overlap (SOE) method. BioTechniques. 1995 Aug;19(2):186–8. 8527132

66. Beyhan S, Bilecen K, Salama SR, Casper-Lindley C, Yildiz FH. 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. 2007 Jan;189(2):388–402. 17071756

67. Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 2001 May 1;29(9):e45. 11328886

68. Robinson MD, McCarthy DJ, Smyth GK. edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics. 2010 Jan 1;26(1):139–40. doi: 10.1093/bioinformatics/btp616 19910308

69. Berk V, Fong JC, Dempsey GT, Develioglu ON, Zhuang X, Liphardt J, et al. Molecular architecture and assembly principles of Vibrio cholerae biofilms. Science. 2012 Jul 13;337(6091):236–9. doi: 10.1126/science.1222981 22798614

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

Článok vyšiel v časopise

PLOS Pathogens


2015 Číslo 5
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Získaná hemofilie - Povědomí o nemoci a její diagnostika
nový kurz

Eozinofilní granulomatóza s polyangiitidou
Autori: doc. MUDr. Martina Doubková, Ph.D.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#