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. 1995 Jul;177(13):3801–3807. doi: 10.1128/jb.177.13.3801-3807.1995

Identification of a Bordetella pertussis regulatory factor required for transcription of the pertussis toxin operon in Escherichia coli.

D DeShazer 1, G E Wood 1, R L Friedman 1
PMCID: PMC177099  PMID: 7601846

Abstract

Transcription of the pertussis toxin operon (ptx) is positively regulated in Bordetella pertussis by the bvgAS locus. However, a ptx-lacZ transcriptional fusion in Escherichia coli cannot be activated by bvgAS in trans. This suggests that an additional factor(s) is required for transcription of ptx. A gene encoding a Bvg accessory factor (Baf) was identified by its ability to activate an E. coli ptx-lacZ fusion in the presence of bvgAS. The expression of ptx-lacZ was decreased by the addition of 40 mM MgSO4, a compound that also modulates ptx expression in B. pertussis. Baf alone did not activate expression of an E. coli fhaB-lacZ fusion, nor did it increase expression of fhaB-lacZ in trans with bvgAS. The gene encoding Baf was localized, sequenced, and found to produce a novel 28-kDa protein. Sequences homologous to B. pertussis baf were identified in Bordetella bronchiseptica and Bordetella parapertussis but not in Bordetella avium. When an additional copy of baf was integrated into the chromosome of BC75, a B. pertussis mutant that produces a low level of pertussis toxin, pertussis toxin production was partially complemented in the cointegrate strain.

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Selected References

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  1. Bannan J. D., Moran M. J., MacInnes J. I., Soltes G. A., Friedman R. L. Cloning and characterization of btr, a Bordetella pertussis gene encoding an FNR-like transcriptional regulator. J Bacteriol. 1993 Nov;175(22):7228–7235. doi: 10.1128/jb.175.22.7228-7235.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boucher P. E., Menozzi F. D., Locht C. The modular architecture of bacterial response regulators. Insights into the activation mechanism of the BvgA transactivator of Bordetella pertussis. J Mol Biol. 1994 Aug 19;241(3):363–377. doi: 10.1006/jmbi.1994.1513. [DOI] [PubMed] [Google Scholar]
  3. Carbonetti N. H., Fuchs T. M., Patamawenu A. A., Irish T. J., Deppisch H., Gross R. Effect of mutations causing overexpression of RNA polymerase alpha subunit on regulation of virulence factors in Bordetella pertussis. J Bacteriol. 1994 Dec;176(23):7267–7273. doi: 10.1128/jb.176.23.7267-7273.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carbonetti N. H., Khelef N., Guiso N., Gross R. A phase variant of Bordetella pertussis with a mutation in a new locus involved in the regulation of pertussis toxin and adenylate cyclase toxin expression. J Bacteriol. 1993 Oct;175(20):6679–6688. doi: 10.1128/jb.175.20.6679-6688.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Charles I. G., Dougan G., Pickard D., Chatfield S., Smith M., Novotny P., Morrissey P., Fairweather N. F. Molecular cloning and characterization of protective outer membrane protein P.69 from Bordetella pertussis. Proc Natl Acad Sci U S A. 1989 May;86(10):3554–3558. doi: 10.1073/pnas.86.10.3554. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cookson B. T., Berg D. E., Goldman W. E. Mutagenesis of Bordetella pertussis with transposon Tn5tac1: conditional expression of virulence-associated genes. J Bacteriol. 1990 Apr;172(4):1681–1687. doi: 10.1128/jb.172.4.1681-1687.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Coote J. G. Antigenic switching and pathogenicity: environmental effects on virulence gene expression in Bordetella pertussis. J Gen Microbiol. 1991 Nov;137(11):2493–2503. doi: 10.1099/00221287-137-11-2493. [DOI] [PubMed] [Google Scholar]
  9. DeShazer D., Bannan J. D., Moran M. J., Friedman R. L. Characterization of the gene encoding superoxide dismutase of Bordetella pertussis and construction of a SOD-deficient mutant. Gene. 1994 May 3;142(1):85–89. doi: 10.1016/0378-1119(94)90359-x. [DOI] [PubMed] [Google Scholar]
  10. DeShazer D., Wood G. E., Friedman R. L. Boiling eliminates artifact banding when sequencing double-stranded templates. Biotechniques. 1994 Aug;17(2):288–290. [PubMed] [Google Scholar]
  11. DeShazer D., Wood G. E., Friedman R. L. Molecular characterization of catalase from Bordetella pertussis: identification of the katA promoter in an upstream insertion sequence. Mol Microbiol. 1994 Oct;14(1):123–130. doi: 10.1111/j.1365-2958.1994.tb01272.x. [DOI] [PubMed] [Google Scholar]
  12. Friedman R. L., Paulaitis S., McMillan J. W. Development of a rapid diagnostic test for pertussis: direct detection of pertussis toxin in respiratory secretions. J Clin Microbiol. 1989 Nov;27(11):2466–2470. doi: 10.1128/jcm.27.11.2466-2470.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Goyard S., Ullmann A. Analysis of Bordetella pertussis cya operon regulation by use of cya-lac fusions. FEMS Microbiol Lett. 1991 Jan 15;61(2-3):251–256. doi: 10.1016/0378-1097(91)90561-n. [DOI] [PubMed] [Google Scholar]
  14. Gross R., Aricò B., Rappuoli R. Families of bacterial signal-transducing proteins. Mol Microbiol. 1989 Nov;3(11):1661–1667. doi: 10.1111/j.1365-2958.1989.tb00152.x. [DOI] [PubMed] [Google Scholar]
  15. Hewlett E. L., Sauer K. T., Myers G. A., Cowell J. L., Guerrant R. L. Induction of a novel morphological response in Chinese hamster ovary cells by pertussis toxin. Infect Immun. 1983 Jun;40(3):1198–1203. doi: 10.1128/iai.40.3.1198-1203.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hohn B., Collins J. A small cosmid for efficient cloning of large DNA fragments. Gene. 1980 Nov;11(3-4):291–298. doi: 10.1016/0378-1119(80)90069-4. [DOI] [PubMed] [Google Scholar]
  17. Ishihama A. Role of the RNA polymerase alpha subunit in transcription activation. Mol Microbiol. 1992 Nov;6(22):3283–3288. doi: 10.1111/j.1365-2958.1992.tb02196.x. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Kuhl S. A., McCreary R. P., Bannan J. D., Friedman R. L. Isolation and characterization of the recA gene of Bordetella pertussis. Mol Microbiol. 1990 Jul;4(7):1165–1172. doi: 10.1111/j.1365-2958.1990.tb00691.x. [DOI] [PubMed] [Google Scholar]
  20. LACEY B. W. Antigenic modulation of Bordetella pertussis. J Hyg (Lond) 1960 Mar;58:57–93. doi: 10.1017/s0022172400038134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Laoide B. M., Ullmann A. Virulence dependent and independent regulation of the Bordetella pertussis cya operon. EMBO J. 1990 Apr;9(4):999–1005. doi: 10.1002/j.1460-2075.1990.tb08202.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Locht C., Keith J. M. Pertussis toxin gene: nucleotide sequence and genetic organization. Science. 1986 Jun 6;232(4755):1258–1264. doi: 10.1126/science.3704651. [DOI] [PubMed] [Google Scholar]
  23. Maskell D. J., Morrissey P., Dougan G. Cloning and nucleotide sequence of the aroA gene of Bordetella pertussis. J Bacteriol. 1988 Jun;170(6):2467–2471. doi: 10.1128/jb.170.6.2467-2471.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Melton A. R., Weiss A. A. Characterization of environmental regulators of Bordetella pertussis. Infect Immun. 1993 Mar;61(3):807–815. doi: 10.1128/iai.61.3.807-815.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Melton A. R., Weiss A. A. Environmental regulation of expression of virulence determinants in Bordetella pertussis. J Bacteriol. 1989 Nov;171(11):6206–6212. doi: 10.1128/jb.171.11.6206-6212.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Miller J. F., Johnson S. A., Black W. J., Beattie D. T., Mekalanos J. J., Falkow S. Constitutive sensory transduction mutations in the Bordetella pertussis bvgS gene. J Bacteriol. 1992 Feb;174(3):970–979. doi: 10.1128/jb.174.3.970-979.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Miller J. F., Mekalanos J. J., Falkow S. Coordinate regulation and sensory transduction in the control of bacterial virulence. Science. 1989 Feb 17;243(4893):916–922. doi: 10.1126/science.2537530. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Nicosia A., Perugini M., Franzini C., Casagli M. C., Borri M. G., Antoni G., Almoni M., Neri P., Ratti G., Rappuoli R. Cloning and sequencing of the pertussis toxin genes: operon structure and gene duplication. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4631–4635. doi: 10.1073/pnas.83.13.4631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nicosia A., Rappuoli R. Promoter of the pertussis toxin operon and production of pertussis toxin. J Bacteriol. 1987 Jun;169(6):2843–2846. doi: 10.1128/jb.169.6.2843-2846.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ogasawara N., Nakai S., Yoshikawa H. Systematic sequencing of the 180 kilobase region of the Bacillus subtilis chromosome containing the replication origin. DNA Res. 1994;1(1):1–14. doi: 10.1093/dnares/1.1.1. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Roy C. R., Miller J. F., Falkow S. Autogenous regulation of the Bordetella pertussis bvgABC operon. Proc Natl Acad Sci U S A. 1990 May;87(10):3763–3767. doi: 10.1073/pnas.87.10.3763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Roy C. R., Miller J. F., Falkow S. The bvgA gene of Bordetella pertussis encodes a transcriptional activator required for coordinate regulation of several virulence genes. J Bacteriol. 1989 Nov;171(11):6338–6344. doi: 10.1128/jb.171.11.6338-6344.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Scarlato V., Aricò B., Rappuoli R. DNA topology affects transcriptional regulation of the pertussis toxin gene of Bordetella pertussis in Escherichia coli and in vitro. J Bacteriol. 1993 Aug;175(15):4764–4771. doi: 10.1128/jb.175.15.4764-4771.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Scarlato V., Aricó B., Domenighini M., Rappuoli R. Environmental regulation of virulence factors in Bordetella species. Bioessays. 1993 Feb;15(2):99–104. doi: 10.1002/bies.950150205. [DOI] [PubMed] [Google Scholar]
  37. Stibitz S., Black W., Falkow S. The construction of a cloning vector designed for gene replacement in Bordetella pertussis. Gene. 1986;50(1-3):133–140. doi: 10.1016/0378-1119(86)90318-5. [DOI] [PubMed] [Google Scholar]
  38. 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]
  39. Stibitz S. Mutations in the bvgA gene of Bordetella pertussis that differentially affect regulation of virulence determinants. J Bacteriol. 1994 Sep;176(18):5615–5621. doi: 10.1128/jb.176.18.5615-5621.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. 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]
  41. 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]
  42. Uhl M. A., Miller J. F. Autophosphorylation and phosphotransfer in the Bordetella pertussis BvgAS signal transduction cascade. Proc Natl Acad Sci U S A. 1994 Feb 1;91(3):1163–1167. doi: 10.1073/pnas.91.3.1163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Weiss A. A., Hewlett E. L., Myers G. A., Falkow S. Tn5-induced mutations affecting virulence factors of Bordetella pertussis. Infect Immun. 1983 Oct;42(1):33–41. doi: 10.1128/iai.42.1.33-41.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. 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]
  45. 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]
  46. Willems R., Paul A., van der Heide H. G., ter Avest A. R., Mooi F. R. Fimbrial phase variation in Bordetella pertussis: a novel mechanism for transcriptional regulation. EMBO J. 1990 Sep;9(9):2803–2809. doi: 10.1002/j.1460-2075.1990.tb07468.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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