Skip to main content
Infection and Immunity logoLink to Infection and Immunity
. 1996 Jan;64(1):283–289. doi: 10.1128/iai.64.1.283-289.1996

Vibrio cholerae Hcp, a secreted protein coregulated with HlyA.

S G Williams 1, L T Varcoe 1, S R Attridge 1, P A Manning 1
PMCID: PMC173757  PMID: 8557353

Abstract

Hcp is a 28-kDa secreted protein of Vibrio cholerae regulated coordinately with the hemolysin, HlyA. Both proteins show a dependence on HlyU for expression, suggesting that Hcp may be secreted by V. cholerae in vivo. We have identified and sequenced two genes for Hcp, designated hcpA and hcpB (hemolysin-coregulated protein). The genes encode identical amino acid sequences. Both express a 28-kDa protein, despite open reading frames with only a 19-kDa capacity, suggesting that the Hcp protein runs aberrantly on polyacrylamide gel electrophoresis. There is no cleavage involved in secretion of Hcp from the cell, suggesting a novel mechanism of secretion. An hcp null mutant was constructed, and this strain displayed no deficiency in virulence or colonization in the infant mouse cholera model. From sequence data and primer extension analysis, we predict that the hcp promoter is the sigma 54 type, with a candidate integration host factor binding site upstream. Although hcp and hlyA are coregulated by HlyU, there are no obvious similarities between their promoters. We predict that an intermediate regulator may be involved in the activation of hcp by HlyU. This raises the possibility that HlyU is part of a regulatory cascade.

Full Text

The Full Text of this article is available as a PDF (439.5 KB).

Selected References

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

  1. Aiba H., Adhya S., de Crombrugghe B. Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem. 1981 Nov 25;256(22):11905–11910. [PubMed] [Google Scholar]
  2. Attridge S. R., Rowley D. The role of the flagellum in the adherence of Vibrio cholerae. J Infect Dis. 1983 May;147(5):864–872. doi: 10.1093/infdis/147.5.864. [DOI] [PubMed] [Google Scholar]
  3. Butterton J. R., Boyko S. A., Calderwood S. B. Use of the Vibrio cholerae irgA gene as a locus for insertion and expression of heterologous antigens in cholera vaccine strains. Vaccine. 1993 Oct;11(13):1327–1335. doi: 10.1016/0264-410x(93)90103-5. [DOI] [PubMed] [Google Scholar]
  4. DiRita V. J. Co-ordinate expression of virulence genes by ToxR in Vibrio cholerae. Mol Microbiol. 1992 Feb;6(4):451–458. doi: 10.1111/j.1365-2958.1992.tb01489.x. [DOI] [PubMed] [Google Scholar]
  5. DiRita V. J. Multiple regulatory systems in Vibrio cholerae pathogenesis. Trends Microbiol. 1994 Feb;2(2):37–38. doi: 10.1016/0966-842x(94)90122-8. [DOI] [PubMed] [Google Scholar]
  6. Donnenberg M. S., Kaper J. B. Construction of an eae deletion mutant of enteropathogenic Escherichia coli by using a positive-selection suicide vector. Infect Immun. 1991 Dec;59(12):4310–4317. doi: 10.1128/iai.59.12.4310-4317.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Economou A., Hamilton W. D., Johnston A. W., Downie J. A. The Rhizobium nodulation gene nodO encodes a Ca2(+)-binding protein that is exported without N-terminal cleavage and is homologous to haemolysin and related proteins. EMBO J. 1990 Feb;9(2):349–354. doi: 10.1002/j.1460-2075.1990.tb08117.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Franzon V. L., Barker A., Manning P. A. Nucleotide sequence encoding the mannose-fucose-resistant hemagglutinin of Vibrio cholerae O1 and construction of a mutant. Infect Immun. 1993 Jul;61(7):3032–3037. doi: 10.1128/iai.61.7.3032-3037.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Freundlich M., Ramani N., Mathew E., Sirko A., Tsui P. The role of integration host factor in gene expression in Escherichia coli. Mol Microbiol. 1992 Sep;6(18):2557–2563. doi: 10.1111/j.1365-2958.1992.tb01432.x. [DOI] [PubMed] [Google Scholar]
  10. Goldberg M. B., Boyko S. A., Calderwood S. B. Positive transcriptional regulation of an iron-regulated virulence gene in Vibrio cholerae. Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1125–1129. doi: 10.1073/pnas.88.4.1125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Issartel J. P., Koronakis V., Hughes C. Activation of Escherichia coli prohaemolysin to the mature toxin by acyl carrier protein-dependent fatty acylation. Nature. 1991 Jun 27;351(6329):759–761. doi: 10.1038/351759a0. [DOI] [PubMed] [Google Scholar]
  13. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  14. Litwin C. M., Calderwood S. B. Analysis of the complexity of gene regulation by fur in Vibrio cholerae. J Bacteriol. 1994 Jan;176(1):240–248. doi: 10.1128/jb.176.1.240-248.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lugtenberg B., Meijers J., Peters R., van der Hoek P., van Alphen L. Electrophoretic resolution of the "major outer membrane protein" of Escherichia coli K12 into four bands. FEBS Lett. 1975 Oct 15;58(1):254–258. doi: 10.1016/0014-5793(75)80272-9. [DOI] [PubMed] [Google Scholar]
  16. Magee A. I., Courtneidge S. A. Two classes of fatty acid acylated proteins exist in eukaryotic cells. EMBO J. 1985 May;4(5):1137–1144. doi: 10.1002/j.1460-2075.1985.tb03751.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Merrick M. J. In a class of its own--the RNA polymerase sigma factor sigma 54 (sigma N). Mol Microbiol. 1993 Dec;10(5):903–909. doi: 10.1111/j.1365-2958.1993.tb00961.x. [DOI] [PubMed] [Google Scholar]
  18. Overbye L. J., Sandkvist M., Bagdasarian M. Genes required for extracellular secretion of enterotoxin are clustered in Vibrio cholerae. Gene. 1993 Sep 30;132(1):101–106. doi: 10.1016/0378-1119(93)90520-d. [DOI] [PubMed] [Google Scholar]
  19. Stoebner J. A., Payne S. M. Iron-regulated hemolysin production and utilization of heme and hemoglobin by Vibrio cholerae. Infect Immun. 1988 Nov;56(11):2891–2895. doi: 10.1128/iai.56.11.2891-2895.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tabor S., Richardson C. C. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074–1078. doi: 10.1073/pnas.82.4.1074. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Thöny B., Hennecke H. The -24/-12 promoter comes of age. FEMS Microbiol Rev. 1989 Dec;5(4):341–357. doi: 10.1016/0168-6445(89)90028-4. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Williams S. G., Attridge S. R., Manning P. A. The transcriptional activator HlyU of Vibrio cholerae: nucleotide sequence and role in virulence gene expression. Mol Microbiol. 1993 Aug;9(4):751–760. doi: 10.1111/j.1365-2958.1993.tb01735.x. [DOI] [PubMed] [Google Scholar]
  24. Williams S. G., Manning P. A. Transcription of the Vibrio cholerae haemolysin gene, hlyA, and cloning of a positive regulatory locus, hlyU. Mol Microbiol. 1991 Aug;5(8):2031–2038. doi: 10.1111/j.1365-2958.1991.tb00825.x. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES