Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1991 Mar 1;88(5):1641–1645. doi: 10.1073/pnas.88.5.1641

ToxR regulates the production of lipoproteins and the expression of serum resistance in Vibrio cholerae.

C Parsot 1, E Taxman 1, J J Mekalanos 1
PMCID: PMC51080  PMID: 2000374

Abstract

The genes encoding three lipoproteins of Vibrio cholerae were identified by a combination of DNA sequence analysis and [3H]palmitate labeling of hybrid proteins encoded by TnphoA gene fusions. The expression of these three lipoproteins, TagA, AcfD, and TcpC, was controlled by ToxR, the cholera toxin transcriptional activator. The involvement of other bacterial lipoproteins in conferring resistance to the bactericidal effects of complement prompted us to examine this possibility in V. cholerae. Remarkably, mutations in toxR and tcp genes (including tcpC), involved in the biogenesis of the toxin coregulated pili, rendered V. cholerae about 10(4)-10(6) times more sensitive to the vibriocidal activity of antibody and complement. Since V. cholerae is a noninvasive organism and toxR and tcp mutants are highly defective in intestinal colonization in animals and humans, these results raise the possibility that resistance to a gut-associated, "complement-like" bactericidal activity may be a major virulence determinant of V. cholerae and other enterobacterial species.

Full text

PDF
1641

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barondess J. J., Beckwith J. A bacterial virulence determinant encoded by lysogenic coliphage lambda. Nature. 1990 Aug 30;346(6287):871–874. doi: 10.1038/346871a0. [DOI] [PubMed] [Google Scholar]
  2. Baselski V. S., Medina R. A., Parker C. D. In vivo and in vitro characterization of virulence-deficient mutants of Vibrio cholerae. Infect Immun. 1979 Apr;24(1):111–116. doi: 10.1128/iai.24.1.111-116.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Binns M. M., Mayden J., Levine R. P. Further characterization of complement resistance conferred on Escherichia coli by the plasmid genes traT of R100 and iss of ColV,I-K94. Infect Immun. 1982 Feb;35(2):654–659. doi: 10.1128/iai.35.2.654-659.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chuba P. J., Leon M. A., Banerjee A., Palchaudhuri S. Cloning and DNA sequence of plasmid determinant iss, coding for increased serum survival and surface exclusion, which has homology with lambda DNA. Mol Gen Genet. 1989 Apr;216(2-3):287–292. doi: 10.1007/BF00334367. [DOI] [PubMed] [Google Scholar]
  5. DiRita V. J., Mekalanos J. J. Genetic regulation of bacterial virulence. Annu Rev Genet. 1989;23:455–482. doi: 10.1146/annurev.ge.23.120189.002323. [DOI] [PubMed] [Google Scholar]
  6. Freter R. Mechanism of Action of Intestinal Antibody in Experimental Cholera II. Antibody-Mediated Antibacterial Reaction at the Mucosal Surface. Infect Immun. 1970 Nov;2(5):556–562. doi: 10.1128/iai.2.5.556-562.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Freter R., O'Brien P. C. Role of chemotaxis in the association of motile bacteria with intestinal mucosa: fitness and virulence of nonchemotactic Vibrio cholerae mutants in infant mice. Infect Immun. 1981 Oct;34(1):222–233. doi: 10.1128/iai.34.1.222-233.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hantke K. Regulation of ferric iron transport in Escherichia coli K12: isolation of a constitutive mutant. Mol Gen Genet. 1981;182(2):288–292. doi: 10.1007/BF00269672. [DOI] [PubMed] [Google Scholar]
  9. Herrington D. A., Hall R. H., Losonsky G., Mekalanos J. J., Taylor R. K., Levine M. M. Toxin, toxin-coregulated pili, and the toxR regulon are essential for Vibrio cholerae pathogenesis in humans. J Exp Med. 1988 Oct 1;168(4):1487–1492. doi: 10.1084/jem.168.4.1487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ichihara S., Hussain M., Mizushima S. Characterization of new membrane lipoproteins and their precursors of Escherichia coli. J Biol Chem. 1981 Mar 25;256(6):3125–3129. [PubMed] [Google Scholar]
  11. Joiner K. A. Complement evasion by bacteria and parasites. Annu Rev Microbiol. 1988;42:201–230. doi: 10.1146/annurev.mi.42.100188.001221. [DOI] [PubMed] [Google Scholar]
  12. Kapperud G., Namork E., Skurnik M., Nesbakken T. Plasmid-mediated surface fibrillae of Yersinia pseudotuberculosis and Yersinia enterocolitica: relationship to the outer membrane protein YOP1 and possible importance for pathogenesis. Infect Immun. 1987 Sep;55(9):2247–2254. doi: 10.1128/iai.55.9.2247-2254.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Lee J. Y., Boman A., Sun C. X., Andersson M., Jörnvall H., Mutt V., Boman H. G. Antibacterial peptides from pig intestine: isolation of a mammalian cecropin. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9159–9162. doi: 10.1073/pnas.86.23.9159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lindberg F., Tennent J. M., Hultgren S. J., Lund B., Normark S. PapD, a periplasmic transport protein in P-pilus biogenesis. J Bacteriol. 1989 Nov;171(11):6052–6058. doi: 10.1128/jb.171.11.6052-6058.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. MEYNELL G. G., SUBBAIAH T. V. Antibacterial mechanisms of the mouse gut. I. Kinetics of infection by Salmonella typhi-murium in normal and streptomycin-treated mice studied with abortive transductants. Br J Exp Pathol. 1963 Apr;44:197–208. [PMC free article] [PubMed] [Google Scholar]
  17. Manoil C., Beckwith J. TnphoA: a transposon probe for protein export signals. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8129–8133. doi: 10.1073/pnas.82.23.8129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mekalanos J. J., Swartz D. J., Pearson G. D., Harford N., Groyne F., de Wilde M. Cholera toxin genes: nucleotide sequence, deletion analysis and vaccine development. Nature. 1983 Dec 8;306(5943):551–557. doi: 10.1038/306551a0. [DOI] [PubMed] [Google Scholar]
  19. Miller V. L., DiRita V. J., Mekalanos J. J. Identification of toxS, a regulatory gene whose product enhances toxR-mediated activation of the cholera toxin promoter. J Bacteriol. 1989 Mar;171(3):1288–1293. doi: 10.1128/jb.171.3.1288-1293.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Miller V. L., Mekalanos J. J. 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. 1988 Jun;170(6):2575–2583. doi: 10.1128/jb.170.6.2575-2583.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Miller V. L., Mekalanos J. J. Synthesis of cholera toxin is positively regulated at the transcriptional level by toxR. Proc Natl Acad Sci U S A. 1984 Jun;81(11):3471–3475. doi: 10.1073/pnas.81.11.3471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Miller V. L., Taylor R. K., Mekalanos J. J. Cholera toxin transcriptional activator toxR is a transmembrane DNA binding protein. Cell. 1987 Jan 30;48(2):271–279. doi: 10.1016/0092-8674(87)90430-2. [DOI] [PubMed] [Google Scholar]
  23. Ogata R. T., Winters C., Levine R. P. Nucleotide sequence analysis of the complement resistance gene from plasmid R100. J Bacteriol. 1982 Aug;151(2):819–827. doi: 10.1128/jb.151.2.819-827.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ouellette A. J., Greco R. M., James M., Frederick D., Naftilan J., Fallon J. T. Developmental regulation of cryptdin, a corticostatin/defensin precursor mRNA in mouse small intestinal crypt epithelium. J Cell Biol. 1989 May;108(5):1687–1695. doi: 10.1083/jcb.108.5.1687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Perumal N. B., Minkley E. G., Jr The product of the F sex factor traT surface exclusion gene is a lipoprotein. J Biol Chem. 1984 May 10;259(9):5357–5360. [PubMed] [Google Scholar]
  26. Peterson K. M., Mekalanos J. J. Characterization of the Vibrio cholerae ToxR regulon: identification of novel genes involved in intestinal colonization. Infect Immun. 1988 Nov;56(11):2822–2829. doi: 10.1128/iai.56.11.2822-2829.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rhen M., Sukupolvi S. The role of the traT gene of the Salmonella typhimurium virulence plasmid for serum resistance and growth within liver macrophages. Microb Pathog. 1988 Oct;5(4):275–285. doi: 10.1016/0882-4010(88)90100-3. [DOI] [PubMed] [Google Scholar]
  28. Shinkai Y., Takio K., Okumura K. Homology of perforin to the ninth component of complement (C9). Nature. 1988 Aug 11;334(6182):525–527. doi: 10.1038/334525a0. [DOI] [PubMed] [Google Scholar]
  29. Sun D. X., Mekalanos J. J., Taylor R. K. Antibodies directed against the toxin-coregulated pilus isolated from Vibrio cholerae provide protection in the infant mouse experimental cholera model. J Infect Dis. 1990 Jun;161(6):1231–1236. doi: 10.1093/infdis/161.6.1231. [DOI] [PubMed] [Google Scholar]
  30. Tagliabue A., Nencioni L., Villa L., Keren D. F., Lowell G. H., Boraschi D. Antibody-dependent cell-mediated antibacterial activity of intestinal lymphocytes with secretory IgA. Nature. 1983 Nov 10;306(5939):184–186. doi: 10.1038/306184a0. [DOI] [PubMed] [Google Scholar]
  31. Taylor R. K., Miller V. L., Furlong D. B., Mekalanos J. J. 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):2833–2837. doi: 10.1073/pnas.84.9.2833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Taylor R., Shaw C., Peterson K., Spears P., Mekalanos J. Safe, live Vibrio cholerae vaccines? Vaccine. 1988 Apr;6(2):151–154. doi: 10.1016/s0264-410x(88)80019-7. [DOI] [PubMed] [Google Scholar]
  33. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wu H. C., Tokunaga M. Biogenesis of lipoproteins in bacteria. Curr Top Microbiol Immunol. 1986;125:127–157. doi: 10.1007/978-3-642-71251-7_9. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES