1887

Abstract

Campylobacter jejuni is an important human pathogen that causes 96 million cases of acute diarrheal disease worldwide each year. We have shown that C. jejuni CsrA is involved in the post-transcriptional regulation of more than 100 proteins, and altered expression of these proteins is presumably involved in the altered virulence-related phenotypes of a csrA mutant. Mutation of fliW results in C. jejuni cells that have greatly truncated flagella, are less motile, less able to form biofilms, and exhibit a reduced ability to colonize chicks. The loss of FliW results in the altered expression of 153 flagellar and non-flagellar proteins, the majority of which are members of the CsrA regulon. The number of proteins dysregulated in the fliW mutant was greater at mid-log phase (120 proteins) than at stationary phase (85 proteins); 52 proteins showed altered expression at both growth phases. Loss of FliW altered the growth-phase- and CsrA-mediated regulation of FlaA flagellin. FliW exerts these effects by binding to both FlaA and to CsrA, as evidenced by pull-down assays, protein-protein cross-linking, and size-exclusion chromatography. Taken together, these results show that CsrA-mediated regulation of both flagellar and non-flagellar proteins is modulated by direct binding of CsrA to the flagellar chaperone FliW. Changing FliW:CsrA stoichiometries at different growth phases allow C. jejuni to couple the expression of flagellar motility to metabolic and virulence characteristics.

Keyword(s): biofilm , motility and protein regulation
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2018-08-16
2024-03-28
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References

  1. WHO WHO Estimates of the Global Burden of Foodborne Diseases: Foodborne Disease Burden Epidemiology Reference Group 2007–2015 Publications of the World Health Organization; 2015
    [Google Scholar]
  2. Troeger C, Forouzanfar M, Rao PC, Khalil I, Brown A. et al. Estimates of global, regional, and national morbidity, mortality, and aetiologies of diarrhoeal diseases: a systematic analysis for the Global Burden of Disease Study 2015. Lancet Infect Dis 2017; 17:909–948 [View Article][PubMed]
    [Google Scholar]
  3. Nachamkin I, Allos BM, Tw H. Campylobacter jejuni infection and the association with Guillain-Barré Syndrome. In Nachamkin I, Blaser MJ. (editors) Campylobacter, 2nd ed. Washington, DC: ASM Press; 2000 pp. 155–175
    [Google Scholar]
  4. Young KT, Davis LM, Dirita VJ. Campylobacter jejuni: molecular biology and pathogenesis. Nat Rev Microbiol 2007; 5:665–679 [View Article][PubMed]
    [Google Scholar]
  5. Haddock G, Mullin M, MacCallum A, Sherry A, Tetley L et al. Campylobacter jejuni 81-176 forms distinct microcolonies on in vitro-infected human small intestinal tissue prior to biofilm formation. Microbiology 2010; 156:3079–3084 [View Article][PubMed]
    [Google Scholar]
  6. Teh KH, Flint S, French N. Biofilm formation by Campylobacter jejuni in controlled mixed-microbial populations. Int J Food Microbiol 2010; 143:118–124 [View Article][PubMed]
    [Google Scholar]
  7. Bronowski C, James CE, Winstanley C. Role of environmental survival in transmission of Campylobacter jejuni. FEMS Microbiol Lett 2014; 356:8–19 [View Article][PubMed]
    [Google Scholar]
  8. Wingender J, Flemming HC. Biofilms in drinking water and their role as reservoir for pathogens. Int J Hyg Environ Health 2011; 214:417–423 [View Article][PubMed]
    [Google Scholar]
  9. Bronowski C, Mustafa K, Goodhead I, James CE, Nelson C et al. Campylobacter jejuni transcriptome changes during loss of culturability in water. PLoS One 2017; 12:e0188936 [View Article][PubMed]
    [Google Scholar]
  10. Fields JA, Li J, Gulbronson CJ, Hendrixson DR, Thompson SA. Campylobacter jejuni CsrA regulates metabolic and virulence associated proteins and is necessary for mouse colonization. PLoS One 2016; 11:e0156932 [View Article][PubMed]
    [Google Scholar]
  11. Fields JA, Thompson SA. Campylobacter jejuni CsrA mediates oxidative stress responses, biofilm formation, and host cell invasion. J Bacteriol 2008; 190:3411–3416 [View Article][PubMed]
    [Google Scholar]
  12. Handley RA, Mulholland F, Reuter M, Ramachandran VK, Musk H et al. PerR controls oxidative stress defence and aerotolerance but not motility-associated phenotypes of Campylobacter jejuni. Microbiology 2015; 161:1524–1536 [View Article][PubMed]
    [Google Scholar]
  13. Kim JC, Oh E, Kim J, Jeon B. Regulation of oxidative stress resistance in Campylobacter jejuni, a microaerophilic foodborne pathogen. Front Microbiol 2015; 6:751 [View Article][PubMed]
    [Google Scholar]
  14. Svensson SL, Davis LM, MacKichan JK, Allan BJ, Pajaniappan M et al. The CprS sensor kinase of the zoonotic pathogen Campylobacter jejuni influences biofilm formation and is required for optimal chick colonization. Mol Microbiol 2009; 71:253–272 [View Article][PubMed]
    [Google Scholar]
  15. Parkhill J, Wren BW, Mungall K, Ketley JM, Churcher C et al. The genome sequence of the food-borne pathogen Campylobacter jejuni reveals hypervariable sequences. Nature 2000; 403:665–668 [View Article][PubMed]
    [Google Scholar]
  16. Raphael BH, Pereira S, Flom GA, Zhang Q, Ketley JM et al. The Campylobacter jejuni response regulator, CbrR, modulates sodium deoxycholate resistance and chicken colonization. J Bacteriol 2005; 187:3662–3670 [View Article][PubMed]
    [Google Scholar]
  17. Fields JA, Thompson SA. Campylobacter jejuni CsrA complements an Escherichia coli csrA mutation for the regulation of biofilm formation, motility and cellular morphology but not glycogen accumulation. BMC Microbiol 2012; 12:233 [View Article][PubMed]
    [Google Scholar]
  18. Babitzke P, Romeo T. CsrB sRNA family: sequestration of RNA-binding regulatory proteins. Curr Opin Microbiol 2007; 10:156–163 [View Article][PubMed]
    [Google Scholar]
  19. Mukherjee S, Yakhnin H, Kysela D, Sokoloski J, Babitzke P et al. CsrA-FliW interaction governs flagellin homeostasis and a checkpoint on flagellar morphogenesis in Bacillus subtilis. Mol Microbiol 2011; 82:447–461 [View Article][PubMed]
    [Google Scholar]
  20. Parrish JR, Yu J, Liu G, Hines JA, Chan JE et al. A proteome-wide protein interaction map for Campylobacter jejuni. Genome Biol 2007; 8:R130 [View Article][PubMed]
    [Google Scholar]
  21. Dugar G, Svensson SL, Bischler T, Wäldchen S, Reinhardt R et al. The CsrA-FliW network controls polar localization of the dual-function flagellin mRNA in Campylobacter jejuni. Nat Commun 2016; 7:11667 [View Article][PubMed]
    [Google Scholar]
  22. Radomska KA, Ordoñez SR, Wösten MM, Wagenaar JA, van Putten JP. Feedback control of Campylobacter jejuni flagellin levels through reciprocal binding of FliW to flagellin and the global regulator CsrA. Mol Microbiol 2016; 102:207–220 [View Article][PubMed]
    [Google Scholar]
  23. Black RE, Levine MM, Clements ML, Hughes TP, Blaser MJ. Experimental Campylobacter jejuni infection in humans. J Infect Dis 1988; 157:472–479 [View Article][PubMed]
    [Google Scholar]
  24. Labigne-Roussel A, Harel J, Tompkins L. Gene transfer from Escherichia coli to Campylobacter species: development of shuttle vectors for genetic analysis of Campylobacter jejuni. J Bacteriol 1987; 169:5320–5323 [View Article][PubMed]
    [Google Scholar]
  25. Hendrixson DR, Dirita VJ. Identification of Campylobacter jejuni genes involved in commensal colonization of the chick gastrointestinal tract. Mol Microbiol 2004; 52:471–484 [View Article][PubMed]
    [Google Scholar]
  26. Pajaniappan M, Hall JE, Cawthraw SA, Newell DG, Gaynor EC et al. A temperature-regulated Campylobacter jejuni gluconate dehydrogenase is involved in respiration-dependent energy conservation and chicken colonization. Mol Microbiol 2008; 68:474–491 [View Article][PubMed]
    [Google Scholar]
  27. Svensson SL, Pryjma M, Gaynor EC. Flagella-mediated adhesion and extracellular DNA release contribute to biofilm formation and stress tolerance of Campylobacter jejuni. PLoS One 2014; 9:e106063 [View Article][PubMed]
    [Google Scholar]
  28. Vakulskas CA, Potts AH, Babitzke P, Ahmer BM, Romeo T. Regulation of bacterial virulence by Csr (Rsm) systems. Microbiol Mol Biol Rev 2015; 79:193–224 [View Article][PubMed]
    [Google Scholar]
  29. Hendrixson DR. Regulation of flagellar gene expression and assembly. In Nachamkin I, Szymanski CM, Blaser MJ. (editors) Campylobacter, 3rd ed. Washington, DC: ASM press; 2008 pp. 545–558
    [Google Scholar]
  30. Joshua GW, Guthrie-Irons C, Karlyshev AV, Wren BW. Biofilm formation in Campylobacter jejuni. Microbiology 2006; 152:387–396 [View Article][PubMed]
    [Google Scholar]
  31. Nachamkin I, Yang XH, Stern NJ. Role of Campylobacter jejuni flagella as colonization factors for three-day-old chicks: analysis with flagellar mutants. Appl Environ Microbiol 1993; 59:1269–1273[PubMed]
    [Google Scholar]
  32. Newell DG, McBride H, Dolby JM. Investigations on the role of flagella in the colonization of infant mice with Campylobacter jejuni and attachment of Campylobacter jejuni to human epithelial cell lines. J Hyg 1985; 95:217–227 [View Article][PubMed]
    [Google Scholar]
  33. Wassenaar TM, van der Zeijst BA, Ayling R, Newell DG. Colonization of chicks by motility mutants of Campylobacter jejuni demonstrates the importance of flagellin A expression. J Gen Microbiol 1993; 139:1171–1175 [View Article][PubMed]
    [Google Scholar]
  34. Mercante J, Edwards AN, Dubey AK, Babitzke P, Romeo T. Molecular geometry of CsrA (RsmA) binding to RNA and its implications for regulated expression. J Mol Biol 2009; 392:511–528 [View Article][PubMed]
    [Google Scholar]
  35. Carrillo CD, Taboada E, Nash JH, Lanthier P, Kelly J et al. Genome-wide expression analyses of Campylobacter jejuni NCTC11168 reveals coordinate regulation of motility and virulence by flhA. J Biol Chem 2004; 279:20327–20338 [View Article][PubMed]
    [Google Scholar]
  36. Yao R, Alm RA, Trust TJ, Guerry P. Construction of new Campylobacter cloning vectors and a new mutational cat cassette. Gene 1993; 130:127–130 [View Article][PubMed]
    [Google Scholar]
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