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. 1984 Jan;43(1):263–269. doi: 10.1128/iai.43.1.263-269.1984

Genetic analysis of phase change in Bordetella pertussis.

A A Weiss, S Falkow
PMCID: PMC263420  PMID: 6317569

Abstract

Avirulent-phase derivatives of Bordetella pertussis (those which have simultaneously lost the ability to synthesize several virulence-associated factors) and the genetic mechanism of the phase change were studied. Increased tolerance to erythromycin was shown to be an avirulent-phase marker. By the use of efficiency of plating on erythromycin, the proportion of avirulent-phase (Vir) variants in a virulent-phase (Vir+) population was determined to be between 10(-3) and 10(-6), depending on the strain. We showed that the phase shift is reversible and detected a complete Vir- to Vir+ to Vir- to cycle. In other experiments, hybridization studies with avirulent-phase mutants obtained by Tn5 mutagenesis suggested that a single region located at a unique site in the B. pertussis chromosome controls the phase change. One of the avirulent Tn5 mutants was used as a recipient in a conjugative cross with a virulent-phase donor. All recombinants which had reacquired the virulence-associated factors also lost Tn5, indicating the loss of Tn5 was required to restore the Vir+ phenotype. The Tn5 avirulent-phase mutants behave as if the insertion interrupted the function of a transacting gene product which is required for the expression of the other virulent-phase genes. A model of the molecular basis of the phase regulation is presented.

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Selected References

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

  1. Ashworth L. A., Irons L. I., Dowsett A. B. Antigenic relationship between serotype-specific agglutinogen and fimbriae of Bordetella pertussis. Infect Immun. 1982 Sep;37(3):1278–1281. doi: 10.1128/iai.37.3.1278-1281.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bauer A. W., Kirby W. M., Sherris J. C., Turck M. Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol. 1966 Apr;45(4):493–496. [PubMed] [Google Scholar]
  3. Berg D. E., Weiss A., Crossland L. Polarity of Tn5 insertion mutations in Escherichia coli. J Bacteriol. 1980 May;142(2):439–446. doi: 10.1128/jb.142.2.439-446.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cowell J. L., Hewlett E. L., Manclark C. R. Intracellular localization of the dermonecrotic toxin of Bordetella pertussis. Infect Immun. 1979 Sep;25(3):896–901. doi: 10.1128/iai.25.3.896-901.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Davies J., Smith D. I. Plasmid-determined resistance to antimicrobial agents. Annu Rev Microbiol. 1978;32:469–518. doi: 10.1146/annurev.mi.32.100178.002345. [DOI] [PubMed] [Google Scholar]
  6. Ezzell J. W., Dobrogosz W. J., Kloos W. E., Manclark C. R. Phase-shift markers in the genus Bordetella: loss of cytochrome d-629 in phase IV variants. Microbios. 1981;31(125-126):171–181. [PubMed] [Google Scholar]
  7. 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]
  8. Hewlett E. L., Urban M. A., Manclark C. R., Wolff J. Extracytoplasmic adenylate cyclase of Bordetella pertussis. Proc Natl Acad Sci U S A. 1976 Jun;73(6):1926–1930. doi: 10.1073/pnas.73.6.1926. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Idigbe E. O., Parton R., Wardlaw A. C. Rapidity of antigenic modulation of Bordetella pertussis in modified Hornibrook medium. J Med Microbiol. 1981 Nov;14(4):409–418. doi: 10.1099/00222615-14-4-409. [DOI] [PubMed] [Google Scholar]
  10. KASUGA T., NAKASE Y., UKISHIMA K., TAKATSU K. Studies on Haemophilis pertussis. III. Some properties of each phase of H. pertussis. Kitasato Arch Exp Med. 1954 Sep;27(3):37–47. [PubMed] [Google Scholar]
  11. KASUGA T., NAKASE Y., UKISHIMA K., TAKATSU K. Studies on Haemophilus pertussis. I. Antigen structure of H. pertussis and its phases. Kitasato Arch Exp Med. 1953 Nov;26(2-3):121–133. [PubMed] [Google Scholar]
  12. 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]
  13. Katada T., Ui M. Direct modification of the membrane adenylate cyclase system by islet-activating protein due to ADP-ribosylation of a membrane protein. Proc Natl Acad Sci U S A. 1982 May;79(10):3129–3133. doi: 10.1073/pnas.79.10.3129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Meyer T. F., Mlawer N., So M. Pilus expression in Neisseria gonorrhoeae involves chromosomal rearrangement. Cell. 1982 Aug;30(1):45–52. doi: 10.1016/0092-8674(82)90010-1. [DOI] [PubMed] [Google Scholar]
  15. Peppler M. S. Isolation and characterization of isogenic pairs of domed hemolytic and flat nonhemolytic colony types of Bordetella pertussis. Infect Immun. 1982 Mar;35(3):840–851. doi: 10.1128/iai.35.3.840-851.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Putnoky P., Kiss G. B., Ott I., Kondorosi A. Tn5 carries a streptomycin resistance determinant downstream from the kanamycin resistance gene. Mol Gen Genet. 1983;191(2):288–294. doi: 10.1007/BF00334828. [DOI] [PubMed] [Google Scholar]
  17. Rothstein S. J., Jorgensen R. A., Postle K., Reznikoff W. S. The inverted repeats of Tn5 are functionally different. Cell. 1980 Mar;19(3):795–805. doi: 10.1016/s0092-8674(80)80055-9. [DOI] [PubMed] [Google Scholar]
  18. Tait R. C., Rodriguez R. L., Boyer H. W. Altered tetracycline resistance in pSC101 recombinant plasmids. Mol Gen Genet. 1977 Mar 16;151(3):327–331. doi: 10.1007/BF00268797. [DOI] [PubMed] [Google Scholar]
  19. Weiss A. A., Falkow S. Transposon insertion and subsequent donor formation promoted by Tn501 in Bordetella pertussis. J Bacteriol. 1983 Jan;153(1):304–309. doi: 10.1128/jb.153.1.304-309.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Wolff J., Cook G. H., Goldhammer A. R., Berkowitz S. A. Calmodulin activates prokaryotic adenylate cyclase. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3841–3844. doi: 10.1073/pnas.77.7.3841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Zieg J., Simon M. Analysis of the nucleotide sequence of an invertible controlling element. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4196–4200. doi: 10.1073/pnas.77.7.4196. [DOI] [PMC free article] [PubMed] [Google Scholar]

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