Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1995 May;177(10):2727–2736. doi: 10.1128/jb.177.10.2727-2736.1995

Identification of a locus required for the regulation of bvg-repressed genes in Bordetella pertussis.

T J Merkel 1, S Stibitz 1
PMCID: PMC176943  PMID: 7751282

Abstract

In Bordetella pertussis, the coordinate regulation of virulence factor expression is controlled by the products of the bvgAS locus. In the presence of modulating signals such as MgSO4, nicotinic acid, or reduced temperature, the expression of bvg-activated genes is reduced while the expression of bvg-repressed genes is induced. One model for the regulation of bvg-repressed genes predicts the existence of a repressor protein encoded by a bvg-activated gene. Once activated, the product of this bvg-activated gene would bind to and repress transcription from the bvg-repressed genes. We isolated five genetically independent transposon insertion mutants of B. pertussis that have a phenotype consistent with the knockout of a putative bvg-regulated repressor. These mutants constitutively expressed a vrg6-phoA transcriptional fusion but demonstrated normal bvgAS function. Genomic mapping and DNA sequence analysis of the sites of transposon insertion demonstrated that these mutants define a locus downstream of bvgAS. Introduction of an in-frame, 12-bp insertion within this locus also conferred the mutant phenotype, confirming that the phenotype seen in the transposon mutants is the result of disruption of a distinct gene, which we have designated bvgR, and is not a consequence of polar effects on bvgAS.

Full Text

The Full Text of this article is available as a PDF (582.2 KB).

Selected References

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

  1. Aricó B., Miller J. F., Roy C., Stibitz S., Monack D., Falkow S., Gross R., Rappuoli R. Sequences required for expression of Bordetella pertussis virulence factors share homology with prokaryotic signal transduction proteins. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6671–6675. doi: 10.1073/pnas.86.17.6671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beattie D. T., Knapp S., Mekalanos J. J. Evidence that modulation requires sequences downstream of the promoters of two vir-repressed genes of Bordetella pertussis. J Bacteriol. 1990 Dec;172(12):6997–7004. doi: 10.1128/jb.172.12.6997-7004.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beattie D. T., Mahan M. J., Mekalanos J. J. Repressor binding to a regulatory site in the DNA coding sequence is sufficient to confer transcriptional regulation of the vir-repressed genes (vrg genes) in Bordetella pertussis. J Bacteriol. 1993 Jan;175(2):519–527. doi: 10.1128/jb.175.2.519-527.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Beattie D. T., Shahin R., Mekalanos J. J. A vir-repressed gene of Bordetella pertussis is required for virulence. Infect Immun. 1992 Feb;60(2):571–577. doi: 10.1128/iai.60.2.571-577.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Black W. J., Falkow S. Construction and characterization of Bordetella pertussis toxin mutants. Infect Immun. 1987 Oct;55(10):2465–2470. doi: 10.1128/iai.55.10.2465-2470.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  7. Brickman E., Beckwith J. Analysis of the regulation of Escherichia coli alkaline phosphatase synthesis using deletions and phi80 transducing phages. J Mol Biol. 1975 Aug 5;96(2):307–316. doi: 10.1016/0022-2836(75)90350-2. [DOI] [PubMed] [Google Scholar]
  8. Brown N. L., Misra T. K., Winnie J. N., Schmidt A., Seiff M., Silver S. The nucleotide sequence of the mercuric resistance operons of plasmid R100 and transposon Tn501: further evidence for mer genes which enhance the activity of the mercuric ion detoxification system. Mol Gen Genet. 1986 Jan;202(1):143–151. doi: 10.1007/BF00330531. [DOI] [PubMed] [Google Scholar]
  9. Chang C. N., Kuang W. J., Chen E. Y. Nucleotide sequence of the alkaline phosphatase gene of Escherichia coli. Gene. 1986;44(1):121–125. doi: 10.1016/0378-1119(86)90050-8. [DOI] [PubMed] [Google Scholar]
  10. Ditta G., Stanfield S., Corbin D., Helinski D. R. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347–7351. doi: 10.1073/pnas.77.12.7347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Finn T. M., Shahin R., Mekalanos J. J. Characterization of vir-activated TnphoA gene fusions in Bordetella pertussis. Infect Immun. 1991 Sep;59(9):3273–3279. doi: 10.1128/iai.59.9.3273-3279.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Griffin H. G., Foster T. J., Silver S., Misra T. K. Cloning and DNA sequence of the mercuric- and organomercurial-resistance determinants of plasmid pDU1358. Proc Natl Acad Sci U S A. 1987 May;84(10):3112–3116. doi: 10.1073/pnas.84.10.3112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Herrero M., de Lorenzo V., Timmis K. N. Transposon vectors containing non-antibiotic resistance selection markers for cloning and stable chromosomal insertion of foreign genes in gram-negative bacteria. J Bacteriol. 1990 Nov;172(11):6557–6567. doi: 10.1128/jb.172.11.6557-6567.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. KASUGA T., NAKASE Y., UKISHIMA K., TAKATSU K. Studies on Haemophilus pertussis. V. Relation between the phase of bacilli and the progress of the whooping-cough. Kitasato Arch Exp Med. 1954 Sep;27(3):57–62. [PubMed] [Google Scholar]
  15. Knapp S., Mekalanos J. J. Two trans-acting regulatory genes (vir and mod) control antigenic modulation in Bordetella pertussis. J Bacteriol. 1988 Nov;170(11):5059–5066. doi: 10.1128/jb.170.11.5059-5066.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Krause M., Guiney D. G. Identification of a multimer resolution system involved in stabilization of the Salmonella dublin virulence plasmid pSDL2. J Bacteriol. 1991 Sep;173(18):5754–5762. doi: 10.1128/jb.173.18.5754-5762.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Miller J. F., Roy C. R., Falkow S. Analysis of Bordetella pertussis virulence gene regulation by use of transcriptional fusions in Escherichia coli. J Bacteriol. 1989 Nov;171(11):6345–6348. doi: 10.1128/jb.171.11.6345-6348.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mooi F. R. Virulence factors of Bordetella pertussis. Antonie Van Leeuwenhoek. 1988;54(5):465–474. doi: 10.1007/BF00461865. [DOI] [PubMed] [Google Scholar]
  19. Parkinson J. S., Kofoid E. C. Communication modules in bacterial signaling proteins. Annu Rev Genet. 1992;26:71–112. doi: 10.1146/annurev.ge.26.120192.000443. [DOI] [PubMed] [Google Scholar]
  20. Roy C. R., Falkow S. Identification of Bordetella pertussis regulatory sequences required for transcriptional activation of the fhaB gene and autoregulation of the bvgAS operon. J Bacteriol. 1991 Apr;173(7):2385–2392. doi: 10.1128/jb.173.7.2385-2392.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Scarlato V., Prugnola A., Aricó B., Rappuoli R. Positive transcriptional feedback at the bvg locus controls expression of virulence factors in Bordetella pertussis. Proc Natl Acad Sci U S A. 1990 Sep;87(17):6753–6757. doi: 10.1073/pnas.87.17.6753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Simons R. W., Houman F., Kleckner N. Improved single and multicopy lac-based cloning vectors for protein and operon fusions. Gene. 1987;53(1):85–96. doi: 10.1016/0378-1119(87)90095-3. [DOI] [PubMed] [Google Scholar]
  23. Stibitz S., Carbonetti N. H. Hfr mapping of mutations in Bordetella pertussis that define a genetic locus involved in virulence gene regulation. J Bacteriol. 1994 Dec;176(23):7260–7266. doi: 10.1128/jb.176.23.7260-7266.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Stibitz S., Garletts T. L. Derivation of a physical map of the chromosome of Bordetella pertussis Tohama I. J Bacteriol. 1992 Dec;174(23):7770–7777. doi: 10.1128/jb.174.23.7770-7777.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Stibitz S. Use of conditionally counterselectable suicide vectors for allelic exchange. Methods Enzymol. 1994;235:458–465. doi: 10.1016/0076-6879(94)35161-9. [DOI] [PubMed] [Google Scholar]
  26. Stibitz S., Weiss A. A., Falkow S. Genetic analysis of a region of the Bordetella pertussis chromosome encoding filamentous hemagglutinin and the pleiotropic regulatory locus vir. J Bacteriol. 1988 Jul;170(7):2904–2913. doi: 10.1128/jb.170.7.2904-2913.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Stibitz S., Yang M. S. Subcellular localization and immunological detection of proteins encoded by the vir locus of Bordetella pertussis. J Bacteriol. 1991 Jul;173(14):4288–4296. doi: 10.1128/jb.173.14.4288-4296.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Weiss A. A., Hewlett E. L. Virulence factors of Bordetella pertussis. Annu Rev Microbiol. 1986;40:661–686. doi: 10.1146/annurev.mi.40.100186.003305. [DOI] [PubMed] [Google Scholar]
  29. Weiss A. A., Melton A. R., Walker K. E., Andraos-Selim C., Meidl J. J. Use of the promoter fusion transposon Tn5 lac to identify mutations in Bordetella pertussis vir-regulated genes. Infect Immun. 1989 Sep;57(9):2674–2682. doi: 10.1128/iai.57.9.2674-2682.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. de Lorenzo V., Herrero M., Jakubzik U., Timmis K. N. Mini-Tn5 transposon derivatives for insertion mutagenesis, promoter probing, and chromosomal insertion of cloned DNA in gram-negative eubacteria. J Bacteriol. 1990 Nov;172(11):6568–6572. doi: 10.1128/jb.172.11.6568-6572.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES