1887

Abstract

moves in response to environmental chemical cues using a chemotaxis two-component signal-transduction system. Autoinducer-2 (AI-2) is a quorum-sensing signal produced by the LuxS protein that accumulates in the bacterial environment in a density-dependent manner. We showed previously that a mutant was defective in motility on soft agar plates. Here we report that deletion of the gene resulted in swimming behaviour with a reduced frequency of stops as compared to the wild-type strain. Stopping frequency was restored to wild-type levels by genetic complementation of the mutation or by addition of synthetic 4,5-dihydroxy-2,3-pentanedione (DPD), which cyclizes to form AI-2. Synthetic DPD also increased the frequency of stops in wild-type , similar to the behaviour induced by the known chemorepellent HCl. We found that whereas mutants lacking the chemoreceptor genes or responded to an exogenous source of synthetic DPD, the chemoreceptor mutant was non-responsive to a gradient or uniform distribution of the chemical. Furthermore, a double mutant lacking both and exhibited chemotactic behaviour similar to the single mutant, whereas a double mutant lacking both and the chemotransduction gene behaved like a nonchemotactic single mutant, supporting the model that functions in a signalling pathway downstream of and upstream of . We conclude that perceives LuxS-produced AI-2 as a chemorepellent via the chemoreceptor TlpB.

Funding
This study was supported by the:
  • National Institutes of Health (Award R01 DK075667 and R01 AI050000)
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.049353-0
2011-09-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/157/9/2445.html?itemId=/content/journal/micro/10.1099/mic.0.049353-0&mimeType=html&fmt=ahah

References

  1. Andermann T. M., Chen Y. T., Ottemann K. M. ( 2002). Two predicted chemoreceptors of Helicobacter pylori promote stomach infection. Infect Immun 70:5877–5881 [View Article][PubMed]
    [Google Scholar]
  2. Armitage J. P. ( 1999). Bacterial tactic responses. Adv Microb Physiol 41:229–289 [View Article][PubMed]
    [Google Scholar]
  3. Bansal T., Jesudhasan P., Pillai S., Wood T. K., Jayaraman A. ( 2008). Temporal regulation of enterohemorrhagic Escherichia coli virulence mediated by autoinducer-2. Appl Microbiol Biotechnol 78:811–819 [View Article][PubMed]
    [Google Scholar]
  4. Bik E. M., Eckburg P. B., Gill S. R., Nelson K. E., Purdom E. A., Francois F., Perez-Perez G., Blaser M. J., Relman D. A. ( 2006). Molecular analysis of the bacterial microbiota in the human stomach. Proc Natl Acad Sci U S A 103:732–737 [View Article][PubMed]
    [Google Scholar]
  5. Blaser M. J., Atherton J. C. ( 2004). Helicobacter pylori persistence: biology and disease. J Clin Invest 113:321–333[PubMed] [CrossRef]
    [Google Scholar]
  6. Boyd A., Simon M. ( 1982). Bacterial chemotaxis. Annu Rev Physiol 44:501–517 [View Article][PubMed]
    [Google Scholar]
  7. Castillo A. R., Arevalo S. S., Woodruff A. J., Ottemann K. M. ( 2008). Experimental analysis of Helicobacter pylori transcriptional terminators suggests this microbe uses both intrinsic and factor-dependent termination. Mol Microbiol 67:155–170 [View Article][PubMed]
    [Google Scholar]
  8. Cerda O., Rivas A., Toledo H. ( 2003). Helicobacter pylori strain ATCC700392 encodes a methyl-accepting chemotaxis receptor protein (MCP) for arginine and sodium bicarbonate. FEMS Microbiol Lett 224:175–181 [View Article][PubMed]
    [Google Scholar]
  9. Chen X., Schauder S., Potier N., Van Dorsselaer A., Pelczer I., Bassler B. L., Hughson F. M. ( 2002). Structural identification of a bacterial quorum-sensing signal containing boron. Nature 415:545–549 [View Article][PubMed]
    [Google Scholar]
  10. Cole S. P., Harwood J., Lee R., She R., Guiney D. G. ( 2004). Characterization of monospecies biofilm formation by Helicobacter pylori . J Bacteriol 186:3124–3132 [View Article][PubMed]
    [Google Scholar]
  11. Copass M., Grandi G., Rappuoli R. ( 1997). Introduction of unmarked mutations in the Helicobacter pylori vacA gene with a sucrose sensitivity marker. Infect Immun 65:1949–1952[PubMed]
    [Google Scholar]
  12. Covacci A., Censini S., Bugnoli M., Petracca R., Burroni D., Macchia G., Massone A., Papini E., Xiang Z. et al. ( 1993). Molecular characterization of the 128-kDa immunodominant antigen of Helicobacter pylori associated with cytotoxicity and duodenal ulcer. Proc Natl Acad Sci U S A 90:5791–5795 [View Article][PubMed]
    [Google Scholar]
  13. Croxen M. A., Sisson G., Melano R., Hoffman P. S. ( 2006). The Helicobacter pylori chemotaxis receptor TlpB (HP0103) is required for pH taxis and for colonization of the gastric mucosa. J Bacteriol 188:2656–2665 [View Article][PubMed]
    [Google Scholar]
  14. Doherty N. C., Shen F., Halliday N. M., Barrett D. A., Hardie K. R., Winzer K., Atherton J. C. ( 2010). In Helicobacter pylori, LuxS is a key enzyme in cysteine provision through a reverse transsulfuration pathway. J Bacteriol 192:1184–1192 [View Article][PubMed]
    [Google Scholar]
  15. Eaton K. A., Morgan D. R., Krakowka S. ( 1992). Motility as a factor in the colonisation of gnotobiotic piglets by Helicobacter pylori . J Med Microbiol 37:123–127 [View Article][PubMed]
    [Google Scholar]
  16. Federle M. J., Bassler B. L. ( 2003). Interspecies communication in bacteria. J Clin Invest 112:1291–1299[PubMed] [CrossRef]
    [Google Scholar]
  17. Forsyth M. H., Cover T. L. ( 2000). Intercellular communication in Helicobacter pylori: luxS is essential for the production of an extracellular signaling molecule. Infect Immun 68:3193–3199 [View Article][PubMed]
    [Google Scholar]
  18. Foynes S., Dorrell N., Ward S. J., Stabler R. A., McColm A. A., Rycroft A. N., Wren B. W. ( 2000). Helicobacter pylori possesses two CheY response regulators and a histidine kinase sensor, CheA, which are essential for chemotaxis and colonization of the gastric mucosa. Infect Immun 68:2016–2023 [View Article][PubMed]
    [Google Scholar]
  19. Hegde M., Englert D. L., Schrock S., Cohn W. B., Vogt C., Wood T. K., Manson M. D., Jayaraman A. ( 2011). Chemotaxis to the quorum-sensing signal AI-2 requires the Tsr chemoreceptor and the periplasmic LsrB AI-2-binding protein. J Bacteriol 193:768–773 [View Article][PubMed]
    [Google Scholar]
  20. Holden N. J., Gally D. L. ( 2004). Switches, cross-talk and memory in Escherichia coli adherence. J Med Microbiol 53:585–593 [View Article][PubMed]
    [Google Scholar]
  21. Joyce E. A., Bassler B. L., Wright A. ( 2000). Evidence for a signaling system in Helicobacter pylori: detection of a luxS-encoded autoinducer. J Bacteriol 182:3638–3643 [View Article][PubMed]
    [Google Scholar]
  22. Lee W. K., Ogura K., Loh J. T., Cover T. L., Berg D. E. ( 2006). Quantitative effect of luxS gene inactivation on the fitness of Helicobacter pylori . Appl Environ Microbiol 72:6615–6622 [View Article][PubMed]
    [Google Scholar]
  23. Loh J. T., Forsyth M. H., Cover T. L. ( 2004). Growth phase regulation of flaA expression in Helicobacter pylori is luxS dependent. Infect Immun 72:5506–5510 [View Article][PubMed]
    [Google Scholar]
  24. Lowenthal A. C., Simon C., Fair A. S., Mehmood K., Terry K., Anastasia S., Ottemann K. M. ( 2009). A fixed-time diffusion analysis method determines that the three cheV genes of Helicobacter pylori differentially affect motility. Microbiology 155:1181–1191 [View Article][PubMed]
    [Google Scholar]
  25. McGee D. J., Coker C., Testerman T. L., Harro J. M., Gibson S. V., Mobley H. L. ( 2002). The Helicobacter pylori flbA flagellar biosynthesis and regulatory gene is required for motility and virulence and modulates urease of H. pylori and Proteus mirabilis . J Med Microbiol 51:958–970[PubMed]
    [Google Scholar]
  26. McGee D. J., Langford M. L., Watson E. L., Carter J. E., Chen Y. T., Ottemann K. M. ( 2005). Colonization and inflammation deficiencies in Mongolian gerbils infected by Helicobacter pylori chemotaxis mutants. Infect Immun 73:1820–1827 [View Article][PubMed]
    [Google Scholar]
  27. Ménard R., Sansonetti P. J., Parsot C. ( 1993). Nonpolar mutagenesis of the ipa genes defines IpaB, IpaC, and IpaD as effectors of Shigella flexneri entry into epithelial cells. J Bacteriol 175:5899–5906[PubMed]
    [Google Scholar]
  28. Miller S. T., Xavier K. B., Campagna S. R., Taga M. E., Semmelhack M. F., Bassler B. L., Hughson F. M. ( 2004). Salmonella typhimurium recognizes a chemically distinct form of the bacterial quorum-sensing signal AI-2. Mol Cell 15:677–687 [View Article][PubMed]
    [Google Scholar]
  29. Ng W. L., Bassler B. L. ( 2009). Bacterial quorum-sensing network architectures. Annu Rev Genet 43:197–222 [View Article][PubMed]
    [Google Scholar]
  30. O’Toole P. W., Lane M. C., Porwollik S. ( 2000). Helicobacter pylori motility. Microbes Infect 2:1207–1214 [View Article][PubMed]
    [Google Scholar]
  31. Osaki T., Hanawa T., Manzoku T., Fukuda M., Kawakami H., Suzuki H., Yamaguchi H., Yan X., Taguchi H. et al. ( 2006). Mutation of luxS affects motility and infectivity of Helicobacter pylori in gastric mucosa of a Mongolian gerbil model. J Med Microbiol 55:1477–1485 [View Article][PubMed]
    [Google Scholar]
  32. Ottemann K. M., Lowenthal A. C. ( 2002). Helicobacter pylori uses motility for initial colonization and to attain robust infection. Infect Immun 70:1984–1990 [View Article][PubMed]
    [Google Scholar]
  33. Pittman M. S., Goodwin M., Kelly D. J. ( 2001). Chemotaxis in the human gastric pathogen Helicobacter pylori: different roles for CheW and the three CheV paralogues, and evidence for CheV2 phosphorylation. Microbiology 147:2493–2504[PubMed]
    [Google Scholar]
  34. Rader B. A., Campagna S. R., Semmelhack M. F., Bassler B. L., Guillemin K. ( 2007). The quorum-sensing molecule autoinducer 2 regulates motility and flagellar morphogenesis in Helicobacter pylori . J Bacteriol 189:6109–6117 [View Article][PubMed]
    [Google Scholar]
  35. Rebbapragada A., Johnson M. S., Harding G. P., Zuccarelli A. J., Fletcher H. M., Zhulin I. B., Taylor B. L. ( 1997). The Aer protein and the serine chemoreceptor Tsr independently sense intracellular energy levels and transduce oxygen, redox, and energy signals for Escherichia coli behavior. Proc Natl Acad Sci U S A 94:10541–10546 [View Article][PubMed]
    [Google Scholar]
  36. Schauder S., Shokat K., Surette M. G., Bassler B. L. ( 2001). The LuxS family of bacterial autoinducers: biosynthesis of a novel quorum-sensing signal molecule. Mol Microbiol 41:463–476 [View Article][PubMed]
    [Google Scholar]
  37. Schreiber S., Konradt M., Groll C., Scheid P., Hanauer G., Werling H. O., Josenhans C., Suerbaum S. ( 2004). The spatial orientation of Helicobacter pylori in the gastric mucus. Proc Natl Acad Sci U S A 101:5024–5029 [View Article][PubMed]
    [Google Scholar]
  38. Schweinitzer T., Mizote T., Ishikawa N., Dudnik A., Inatsu S., Schreiber S., Suerbaum S., Aizawa S., Josenhans C. ( 2008). Functional characterization and mutagenesis of the proposed behavioral sensor TlpD of Helicobacter pylori . J Bacteriol 190:3244–3255 [View Article][PubMed]
    [Google Scholar]
  39. Semmelhack M. F., Campagna S. R., Federle M. J., Bassler B. L. ( 2005). An expeditious synthesis of DPD and boron binding studies. Org Lett 7:569–572 [View Article][PubMed]
    [Google Scholar]
  40. Shao H., James D., Lamont R. J., Demuth D. R. ( 2007). Differential interaction of Aggregatibacter (Actinobacillus) actinomycetemcomitans LsrB and RbsB proteins with autoinducer 2. J Bacteriol 189:5559–5565 [View Article][PubMed]
    [Google Scholar]
  41. Shen F., Hobley L., Doherty N., Loh J. T., Cover T. L., Sockett R. E., Hardie K. R., Atherton J. C. ( 2010). In Helicobacter pylori auto-inducer-2, but not LuxS/MccAB catalysed reverse transsulphuration, regulates motility through modulation of flagellar gene transcription. BMC Microbiol 10:210 [View Article][PubMed]
    [Google Scholar]
  42. Szurmant H., Ordal G. W. ( 2004). Diversity in chemotaxis mechanisms among the bacteria and archaea. Microbiol Mol Biol Rev 68:301–319 [View Article][PubMed]
    [Google Scholar]
  43. Terry K., Williams S. M., Connolly L., Ottemann K. M. ( 2005). Chemotaxis plays multiple roles during Helicobacter pylori animal infection. Infect Immun 73:803–811 [View Article][PubMed]
    [Google Scholar]
  44. Terry K., Go A. C., Ottemann K. M. ( 2006). Proteomic mapping of a suppressor of non-chemotactic cheW mutants reveals that Helicobacter pylori contains a new chemotaxis protein. Mol Microbiol 61:871–882 [View Article][PubMed]
    [Google Scholar]
  45. Williams S. M., Chen Y. T., Andermann T. M., Carter J. E., McGee D. J., Ottemann K. M. ( 2007). Helicobacter pylori chemotaxis modulates inflammation and bacterium-gastric epithelium interactions in infected mice. Infect Immun 75:3747–3757 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.049353-0
Loading
/content/journal/micro/10.1099/mic.0.049353-0
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