Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1997 Aug;179(15):4647–4653. doi: 10.1128/jb.179.15.4647-4653.1997

The flgE gene of Campylobacter coli is under the control of the alternative sigma factor sigma54.

N Kinsella 1, P Guerry 1, J Cooney 1, T J Trust 1
PMCID: PMC179307  PMID: 9244248

Abstract

The flgE gene encoding the flagellar hook protein of Campylobacter coli VC167-T1 was cloned by immunoscreening of a genomic library constructed in lambdaZAP Express. The flgE DNA sequence was 2,553 bp in length and encoded a protein with a deduced molecular mass of 90,639 Da. The sequence had significant homology to the 5' and 3' sequences of the flgE genes of Helicobacter pylori, Treponema phagedenis, and Salmonella typhimurium. Primer extension analysis indicated that the VC167 flgE gene is controlled by a sigma54 promoter. PCR analysis showed that the flgE gene size and the 5' and 3' DNA sequences were conserved among C. coli and C. jejuni strains. Southern hybridization analyses confirmed that there is considerable sequence identity among the hook genes of C. coli and C. jejuni but that there are also regions within the genes which differ. Mutants of C. coli defective in hook production were generated by allele replacement. These mutants were nonmotile and lacked flagellar filaments. Analyses of flgE mutants indicated that the carboxy terminus of FlgE is necessary for assembly of the hook structure but not for secretion of FlgE and that, unlike salmonellae, the lack of flgE expression does not result in repression of flagellin expression.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

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

  1. Aizawa S. I., Dean G. E., Jones C. J., Macnab R. M., Yamaguchi S. Purification and characterization of the flagellar hook-basal body complex of Salmonella typhimurium. J Bacteriol. 1985 Mar;161(3):836–849. doi: 10.1128/jb.161.3.836-849.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. 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]
  4. 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]
  5. 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]
  6. Carpenter P. B., Ordal G. W. Bacillus subtilis FlhA: a flagellar protein related to a new family of signal-transducing receptors. Mol Microbiol. 1993 Mar;7(5):735–743. doi: 10.1111/j.1365-2958.1993.tb01164.x. [DOI] [PubMed] [Google Scholar]
  7. Crothers D. M., Haran T. E., Nadeau J. G. Intrinsically bent DNA. J Biol Chem. 1990 May 5;265(13):7093–7096. [PubMed] [Google Scholar]
  8. DePamphilis M. L., Adler J. Fine structure and isolation of the hook-basal body complex of flagella from Escherichia coli and Bacillus subtilis. J Bacteriol. 1971 Jan;105(1):384–395. doi: 10.1128/jb.105.1.384-395.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gillen K. L., Hughes K. T. Molecular characterization of flgM, a gene encoding a negative regulator of flagellin synthesis in Salmonella typhimurium. J Bacteriol. 1991 Oct;173(20):6453–6459. doi: 10.1128/jb.173.20.6453-6459.1991. [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., Doig P., Alm R. A., Burr D. H., Kinsella N., Trust T. J. Identification and characterization of genes required for post-translational modification of Campylobacter coli VC167 flagellin. Mol Microbiol. 1996 Jan;19(2):369–378. doi: 10.1046/j.1365-2958.1996.369895.x. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Gustafson C. E., Thomas C. J., Trust T. J. Detection of Aeromonas salmonicida from fish by using polymerase chain reaction amplification of the virulence surface array protein gene. Appl Environ Microbiol. 1992 Dec;58(12):3816–3825. doi: 10.1128/aem.58.12.3816-3825.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  15. Homma M., DeRosier D. J., Macnab R. M. Flagellar hook and hook-associated proteins of Salmonella typhimurium and their relationship to other axial components of the flagellum. J Mol Biol. 1990 Jun 20;213(4):819–832. doi: 10.1016/S0022-2836(05)80266-9. [DOI] [PubMed] [Google Scholar]
  16. Hughes K. T., Gillen K. L., Semon M. J., Karlinsey J. E. Sensing structural intermediates in bacterial flagellar assembly by export of a negative regulator. Science. 1993 Nov 19;262(5137):1277–1280. doi: 10.1126/science.8235660. [DOI] [PubMed] [Google Scholar]
  17. Jin T., Penner J. L. Role of the 92.5-kilodalton outer membrane protein of Campylobacter jejuni in serological reactions. J Clin Microbiol. 1988 Dec;26(12):2480–2483. doi: 10.1128/jcm.26.12.2480-2483.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Josenhans C., Labigne A., Suerbaum S. Comparative ultrastructural and functional studies of Helicobacter pylori and Helicobacter mustelae flagellin mutants: both flagellin subunits, FlaA and FlaB, are necessary for full motility in Helicobacter species. J Bacteriol. 1995 Jun;177(11):3010–3020. doi: 10.1128/jb.177.11.3010-3020.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kawagishi I., Müller V., Williams A. W., Irikura V. M., Macnab R. M. Subdivision of flagellar region III of the Escherichia coli and Salmonella typhimurium chromosomes and identification of two additional flagellar genes. J Gen Microbiol. 1992 Jun;138(6):1051–1065. doi: 10.1099/00221287-138-6-1051. [DOI] [PubMed] [Google Scholar]
  20. Kornacker M. G., Newton A. Information essential for cell-cycle-dependent secretion of the 591-residue Caulobacter hook protein is confined to a 21-amino-acid sequence near the N-terminus. Mol Microbiol. 1994 Oct;14(1):73–85. doi: 10.1111/j.1365-2958.1994.tb01268.x. [DOI] [PubMed] [Google Scholar]
  21. Kranz R. G., Foster-Hartnett D. Transcriptional regulatory cascade of nitrogen-fixation genes in anoxygenic photosynthetic bacteria: oxygen- and nitrogen-responsive factors. Mol Microbiol. 1990 Nov;4(11):1793–1800. doi: 10.1111/j.1365-2958.1990.tb02027.x. [DOI] [PubMed] [Google Scholar]
  22. Labigne-Roussel A., Courcoux P., Tompkins L. Gene disruption and replacement as a feasible approach for mutagenesis of Campylobacter jejuni. J Bacteriol. 1988 Apr;170(4):1704–1708. doi: 10.1128/jb.170.4.1704-1708.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  24. Lagenaur C., DeMartini M., Agabian N. Isolation and characterization of Caulobacter crescentus flagellar hooks. J Bacteriol. 1978 Nov;136(2):795–798. doi: 10.1128/jb.136.2.795-798.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Limberger R. J., Slivienski L. L., Samsonoff W. A. Genetic and biochemical analysis of the flagellar hook of Treponema phagedenis. J Bacteriol. 1994 Jun;176(12):3631–3637. doi: 10.1128/jb.176.12.3631-3637.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Logan S. M., Guerry P., Rollins D. M., Burr D. H., Trust T. J. In vivo antigenic variation of Campylobacter flagellin. Infect Immun. 1989 Aug;57(8):2583–2585. doi: 10.1128/iai.57.8.2583-2585.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Logan S. M., Trust T. J. Location of epitopes on Campylobacter jejuni flagella. J Bacteriol. 1986 Nov;168(2):739–745. doi: 10.1128/jb.168.2.739-745.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Logan S. M., Trust T. J. Molecular identification of surface protein antigens of Campylobacter jejuni. Infect Immun. 1983 Nov;42(2):675–682. doi: 10.1128/iai.42.2.675-682.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Macnab R. M. Genetics and biogenesis of bacterial flagella. Annu Rev Genet. 1992;26:131–158. doi: 10.1146/annurev.ge.26.120192.001023. [DOI] [PubMed] [Google Scholar]
  30. Miller S., Pesci E. C., Pickett C. L. A Campylobacter jejuni homolog of the LcrD/FlbF family of proteins is necessary for flagellar biogenesis. Infect Immun. 1993 Jul;61(7):2930–2936. doi: 10.1128/iai.61.7.2930-2936.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Miller S., Pesci E. C., Pickett C. L. Genetic organization of the region upstream from the Campylobacter jejuni flagellar gene flhA. Gene. 1994 Aug 19;146(1):31–38. doi: 10.1016/0378-1119(94)90830-3. [DOI] [PubMed] [Google Scholar]
  32. Mills S. D., Bradbury W. C. Human antibody response to outer membrane proteins of Campylobacter jejuni during infection. Infect Immun. 1984 Feb;43(2):739–743. doi: 10.1128/iai.43.2.739-743.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Mirel D. B., Lauer P., Chamberlin M. J. Identification of flagellar synthesis regulatory and structural genes in a sigma D-dependent operon of Bacillus subtilis. J Bacteriol. 1994 Aug;176(15):4492–4500. doi: 10.1128/jb.176.15.4492-4500.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. 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]
  35. Needleman S. B., Wunsch C. D. A general method applicable to the search for similarities in the amino acid sequence of two proteins. J Mol Biol. 1970 Mar;48(3):443–453. doi: 10.1016/0022-2836(70)90057-4. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. 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]
  38. O'Toole P. W., Kostrzynska M., Trust T. J. Non-motile mutants of Helicobacter pylori and Helicobacter mustelae defective in flagellar hook production. Mol Microbiol. 1994 Nov;14(4):691–703. doi: 10.1111/j.1365-2958.1994.tb01307.x. [DOI] [PubMed] [Google Scholar]
  39. Pavlovskis O. R., Rollins D. M., Haberberger R. L., Jr, Green A. E., Habash L., Strocko S., Walker R. I. Significance of flagella in colonization resistance of rabbits immunized with Campylobacter spp. Infect Immun. 1991 Jul;59(7):2259–2264. doi: 10.1128/iai.59.7.2259-2264.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Power M. E., Alm R. A., Trust T. J. Biochemical and antigenic properties of the Campylobacter flagellar hook protein. J Bacteriol. 1992 Jun;174(12):3874–3883. doi: 10.1128/jb.174.12.3874-3883.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Sasse-Dwight S., Gralla J. D. Role of eukaryotic-type functional domains found in the prokaryotic enhancer receptor factor sigma 54. Cell. 1990 Sep 7;62(5):945–954. doi: 10.1016/0092-8674(90)90269-k. [DOI] [PubMed] [Google Scholar]
  42. Schmitz A., Josenhans C., Suerbaum S. Cloning and characterization of the Helicobacter pylori flbA gene, which codes for a membrane protein involved in coordinated expression of flagellar genes. J Bacteriol. 1997 Feb;179(4):987–997. doi: 10.1128/jb.179.4.987-997.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. 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]
  44. Vonderviszt F., Závodszky P., Ishimura M., Uedaira H., Namba K. Structural organization and assembly of flagellar hook protein from Salmonella typhimurium. J Mol Biol. 1995 Aug 25;251(4):520–532. doi: 10.1006/jmbi.1995.0453. [DOI] [PubMed] [Google Scholar]
  45. Wang Y., Taylor D. E. Chloramphenicol resistance in Campylobacter coli: nucleotide sequence, expression, and cloning vector construction. Gene. 1990 Sep 28;94(1):23–28. doi: 10.1016/0378-1119(90)90463-2. [DOI] [PubMed] [Google Scholar]
  46. 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]
  47. Yao R., Burr D. H., Doig P., Trust T. J., Niu H., Guerry P. Isolation of motile and non-motile insertional mutants of Campylobacter jejuni: the role of motility in adherence and invasion of eukaryotic cells. Mol Microbiol. 1994 Dec;14(5):883–893. doi: 10.1111/j.1365-2958.1994.tb01324.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES