Differential flagellin expression in a flaA flaB+ mutant of Campylobacter jejuni (original) (raw)

Abstract

Campylobacter jejuni 81116 has two genes coding for flagellin, flaA and flaB. Fully motile wild-type C. jejuni bacteria express the flaA gene, with no flaB message being detected. A nonmotile flaA flaB+ mutant, R1, produced detectable levels of flagellin B which was incorporated into truncated flagella. After R1 had invaded INT-407 cells, a variant with increased motility, R1-V2, was isolated. R1-V2 produced full-length flagella and an increased amount of flagellin B. Transcriptional analysis showed that R1-V2 contained more flaB mRNA than its parental strain, R1. The flaB gene promoter sequence and primer extension experiments confirmed that transcription of the flaB gene is initiated from a sigma 54 promoter. Neither the promoter sequence nor the coding sequence of flaB had changed in R1-V2. In contrast to R1, R1-V2 no longer produced (truncated) flaA mRNA. The sigma 28 flaA promoter sequence was not changed in R1-V2. We propose that expression of the two flagellin genes in C. jejuni 81116 is regulated at the transcriptional level, in such a way that predominantly one gene at a time is transcribed. We compared the levels of invasiveness of the wild-type strain, R1, and R1-V2 for INT-407 cells. The shift in expression from flaA to flaB occurred not only during invasion assays but also under different conditions in the absence of eukaryotic cells.

3901

Images in this article

Selected References

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

  1. Aguero-Rosenfeld M. E., Yang X. H., Nachamkin I. Infection of adult Syrian hamsters with flagellar variants of Campylobacter jejuni. Infect Immun. 1990 Jul;58(7):2214–2219. doi: 10.1128/iai.58.7.2214-2219.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alm R. A., Guerry P., Power M. E., Trust T. J. Variation in antigenicity and molecular weight of Campylobacter coli VC167 flagellin in different genetic backgrounds. J Bacteriol. 1992 Jul;174(13):4230–4238. doi: 10.1128/jb.174.13.4230-4238.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Alm R. A., Guerry P., Trust T. J. Significance of duplicated flagellin genes in Campylobacter. J Mol Biol. 1993 Mar 20;230(2):359–363. doi: 10.1006/jmbi.1993.1151. [DOI] [PubMed] [Google Scholar]
  4. Alm R. A., Guerry P., Trust T. J. The Campylobacter sigma 54 flaB flagellin promoter is subject to environmental regulation. J Bacteriol. 1993 Jul;175(14):4448–4455. doi: 10.1128/jb.175.14.4448-4455.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Black R. E., Levine M. M., Clements M. L., Hughes T. P., Blaser M. J. Experimental Campylobacter jejuni infection in humans. J Infect Dis. 1988 Mar;157(3):472–479. doi: 10.1093/infdis/157.3.472. [DOI] [PubMed] [Google Scholar]
  6. Caldwell M. B., Guerry P., Lee E. C., Burans J. P., Walker R. I. Reversible expression of flagella in Campylobacter jejuni. Infect Immun. 1985 Dec;50(3):941–943. doi: 10.1128/iai.50.3.941-943.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ferrero R. L., Lee A. Motility of Campylobacter jejuni in a viscous environment: comparison with conventional rod-shaped bacteria. J Gen Microbiol. 1988 Jan;134(1):53–59. doi: 10.1099/00221287-134-1-53. [DOI] [PubMed] [Google Scholar]
  8. Fischer S. H., Nachamkin I. Common and variable domains of the flagellin gene, flaA, in Campylobacter jejuni. Mol Microbiol. 1991 May;5(5):1151–1158. doi: 10.1111/j.1365-2958.1991.tb01888.x. [DOI] [PubMed] [Google Scholar]
  9. Grant C. C., Konkel M. E., Cieplak W., Jr, Tompkins L. S. Role of flagella in adherence, internalization, and translocation of Campylobacter jejuni in nonpolarized and polarized epithelial cell cultures. Infect Immun. 1993 May;61(5):1764–1771. doi: 10.1128/iai.61.5.1764-1771.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Guerry P., Alm R. A., Power M. E., Logan S. M., Trust T. J. Role of two flagellin genes in Campylobacter motility. J Bacteriol. 1991 Aug;173(15):4757–4764. doi: 10.1128/jb.173.15.4757-4764.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Guerry P., Logan S. M., Thornton S., Trust T. J. Genomic organization and expression of Campylobacter flagellin genes. J Bacteriol. 1990 Apr;172(4):1853–1860. doi: 10.1128/jb.172.4.1853-1860.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Harris L. A., Logan S. M., Guerry P., Trust T. J. Antigenic variation of Campylobacter flagella. J Bacteriol. 1987 Nov;169(11):5066–5071. doi: 10.1128/jb.169.11.5066-5071.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Helmann J. D. Alternative sigma factors and the regulation of flagellar gene expression. Mol Microbiol. 1991 Dec;5(12):2875–2882. doi: 10.1111/j.1365-2958.1991.tb01847.x. [DOI] [PubMed] [Google Scholar]
  14. Helmann J. D., Chamberlin M. J. Structure and function of bacterial sigma factors. Annu Rev Biochem. 1988;57:839–872. doi: 10.1146/annurev.bi.57.070188.004203. [DOI] [PubMed] [Google Scholar]
  15. Kanto S., Okino H., Aizawa S., Yamaguchi S. Amino acids responsible for flagellar shape are distributed in terminal regions of flagellin. J Mol Biol. 1991 Jun 5;219(3):471–480. doi: 10.1016/0022-2836(91)90187-b. [DOI] [PubMed] [Google Scholar]
  16. Kustu S., Santero E., Keener J., Popham D., Weiss D. Expression of sigma 54 (ntrA)-dependent genes is probably united by a common mechanism. Microbiol Rev. 1989 Sep;53(3):367–376. doi: 10.1128/mr.53.3.367-376.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Morett E., Buck M. In vivo studies on the interaction of RNA polymerase-sigma 54 with the Klebsiella pneumoniae and Rhizobium meliloti nifH promoters. The role of NifA in the formation of an open promoter complex. J Mol Biol. 1989 Nov 5;210(1):65–77. doi: 10.1016/0022-2836(89)90291-x. [DOI] [PubMed] [Google Scholar]
  18. Morooka T., Umeda A., Amako K. Motility as an intestinal colonization factor for Campylobacter jejuni. J Gen Microbiol. 1985 Aug;131(8):1973–1980. doi: 10.1099/00221287-131-8-1973. [DOI] [PubMed] [Google Scholar]
  19. Newell D. G., McBride H., Dolby J. M. Investigations on the role of flagella in the colonization of infant mice with Campylobacter jejuni and attachment of Campylobacter jejuni to human epithelial cell lines. J Hyg (Lond) 1985 Oct;95(2):217–227. doi: 10.1017/s0022172400062653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Newell D. G. Monoclonal antibodies directed against the flagella of Campylobacter jejuni: production, characterization and lack of effect on the colonization of infant mice. J Hyg (Lond) 1986 Apr;96(2):131–141. doi: 10.1017/s0022172400065906. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ninfa A. J., Mullin D. A., Ramakrishnan G., Newton A. Escherichia coli sigma 54 RNA polymerase recognizes Caulobacter crescentus flbG and flaN flagellar gene promoters in vitro. J Bacteriol. 1989 Jan;171(1):383–391. doi: 10.1128/jb.171.1.383-391.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nuijten P. J., Bleumink-Pluym N. M., Gaastra W., van der Zeijst B. A. Flagellin expression in Campylobacter jejuni is regulated at the transcriptional level. Infect Immun. 1989 Apr;57(4):1084–1088. doi: 10.1128/iai.57.4.1084-1088.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nuijten P. J., van Asten F. J., Gaastra W., van der Zeijst B. A. Structural and functional analysis of two Campylobacter jejuni flagellin genes. J Biol Chem. 1990 Oct 15;265(29):17798–17804. [PubMed] [Google Scholar]
  24. Nuijten P. J., van der Zeijst B. A., Newell D. G. Localization of immunogenic regions on the flagellin proteins of Campylobacter jejuni 81116. Infect Immun. 1991 Mar;59(3):1100–1105. doi: 10.1128/iai.59.3.1100-1105.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Silverman M., Simon M. Phase variation: genetic analysis of switching mutants. Cell. 1980 Apr;19(4):845–854. doi: 10.1016/0092-8674(80)90075-6. [DOI] [PubMed] [Google Scholar]
  26. Totten P. A., Lara J. C., Lory S. The rpoN gene product of Pseudomonas aeruginosa is required for expression of diverse genes, including the flagellin gene. J Bacteriol. 1990 Jan;172(1):389–396. doi: 10.1128/jb.172.1.389-396.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wassenaar T. M., Bleumink-Pluym N. M., van der Zeijst B. A. Inactivation of Campylobacter jejuni flagellin genes by homologous recombination demonstrates that flaA but not flaB is required for invasion. EMBO J. 1991 Aug;10(8):2055–2061. doi: 10.1002/j.1460-2075.1991.tb07736.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wassenaar T. M., van der Zeijst B. A., Ayling R., Newell D. G. Colonization of chicks by motility mutants of Campylobacter jejuni demonstrates the importance of flagellin A expression. J Gen Microbiol. 1993 Jun;139(Pt 6):1171–1175. doi: 10.1099/00221287-139-6-1171. [DOI] [PubMed] [Google Scholar]