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. 1989 Jun;171(6):3031–3038. doi: 10.1128/jb.171.6.3031-3038.1989

Evidence for posttranslational modification and gene duplication of Campylobacter flagellin.

S M Logan 1, T J Trust 1, P Guerry 1
PMCID: PMC210011  PMID: 2722741

Abstract

A gene encoding a flagellin protein of Campylobacter coli VC167 has been cloned and sequenced. The gene was identified in a pBR322 library by hybridization to a synthetic oligonucleotide probe corresponding to amino acids 4 to 9 of the N-terminal sequence obtained by direct chemical analysis (S. M. Logan, L. A. Harris, and T. J. Trust, J. Bacteriol. 169:5072-5077, 1987). The DNA was sequenced and shown to contain an open reading frame encoding a protein with a molecular weight of 58,945 and a length of 572 amino acids. The deduced amino acid sequence was identical to the published N-terminal amino acid sequence of VC167 flagellin and to four internal regions whose partial sequences were obtained by direct chemical analysis of two tryptic and two cyanogen bromide peptides of VC167 flagellin. The C. coli flagellin protein contains posttranslationally modified serine residues, most of which occur within a region containing two 9-amino-acid repeating peptides separated by 34 unique amino acids. Comparisons with the sequences of flagellins from other bacterial species revealed conserved residues at the amino- and carboxy-terminal regions. Hybridization data suggest the presence of a second flagellin copy located adjacent to the first on the VC167 chromosome.

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Selected References

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  1. AMBLER R. P., REES M. W. Epsilon-N-Methyl-lysine in bacterial flagellar protein. Nature. 1959 Jul 4;184:56–57. doi: 10.1038/184056b0. [DOI] [PubMed] [Google Scholar]
  2. Beachey E. H., Seyer J. M., Kang A. H. Repeating covalent structure of streptococcal M protein. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3163–3167. doi: 10.1073/pnas.75.7.3163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Belland R. J., Trust T. J. Deoxyribonucleic acid sequence relatedness between thermophilic members of the genus Campylobacter. J Gen Microbiol. 1982 Nov;128(11):2515–2522. doi: 10.1099/00221287-128-11-2515. [DOI] [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. Butzler J. P., Skirrow M. B. Campylobacter enteritis. Clin Gastroenterol. 1979 Sep;8(3):737–765. [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. Chowdhury M. N. Campylobacter jejuni enteritis; a review. Trop Geogr Med. 1984 Sep;36(3):215–222. [PubMed] [Google Scholar]
  8. Clewell D. B., Helinski D. R. Effect of growth conditions on the formation of the relaxation complex of supercoiled ColE1 deoxyribonucleic acid and protein in Escherichia coli. J Bacteriol. 1972 Jun;110(3):1135–1146. doi: 10.1128/jb.110.3.1135-1146.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. DeLange R. J., Chang J. Y., Shaper J. H., Glazer A. N. Amino acid sequence of flagellin of Bacillus subtilis 168. III. Tryptic peptides, N-bromosuccinimide peptides, and the complete amino acid sequence. J Biol Chem. 1976 Feb 10;251(3):705–711. [PubMed] [Google Scholar]
  10. Froholm L. O., Sletten K. Purification and N-terminal sequence of a fimbrial protein from Moraxella nonliquefaciens. FEBS Lett. 1977 Jan 15;73(1):29–32. doi: 10.1016/0014-5793(77)80008-2. [DOI] [PubMed] [Google Scholar]
  11. Glazer A. N., De Lange R. J., Martinez R. J. Identification of episilon-N-methyllysine in Spirillum serpens flagella and of episilon-N-dimethyllysine in Salmonella typhimurium flagella. Biochim Biophys Acta. 1969 Aug 12;188(1):164–165. doi: 10.1016/0005-2795(69)90059-2. [DOI] [PubMed] [Google Scholar]
  12. Groisman E. A., Casadaban M. J. Mini-mu bacteriophage with plasmid replicons for in vivo cloning and lac gene fusing. J Bacteriol. 1986 Oct;168(1):357–364. doi: 10.1128/jb.168.1.357-364.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Guerry P., Logan S. M., Trust T. J. Genomic rearrangements associated with antigenic variation in Campylobacter coli. J Bacteriol. 1988 Jan;170(1):316–319. doi: 10.1128/jb.170.1.316-319.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Hermodson M. A., Chen K. C., Buchanan T. M. Neisseria pili proteins: amino-terminal amino acid sequences and identification of an unusual amino acid. Biochemistry. 1978 Feb 7;17(3):442–445. doi: 10.1021/bi00596a010. [DOI] [PubMed] [Google Scholar]
  16. Homma M., Fujita H., Yamaguchi S., Iino T. Regions of Salmonella typhimurium flagellin essential for its polymerization and excretion. J Bacteriol. 1987 Jan;169(1):291–296. doi: 10.1128/jb.169.1.291-296.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hull R. A., Gill R. E., Hsu P., Minshew B. H., Falkow S. Construction and expression of recombinant plasmids encoding type 1 or D-mannose-resistant pili from a urinary tract infection Escherichia coli isolate. Infect Immun. 1981 Sep;33(3):933–938. doi: 10.1128/iai.33.3.933-938.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Iino T. Genetics of structure and function of bacterial flagella. Annu Rev Genet. 1977;11:161–182. doi: 10.1146/annurev.ge.11.120177.001113. [DOI] [PubMed] [Google Scholar]
  19. Joys T. M. Identification of an antibody binding site in the phase-1 flagellar protein of Salmonella typhimurium. Microbios. 1976;15(61-62):221–228. [PubMed] [Google Scholar]
  20. Joys T. M., Rankis V. The primary structure of the phase-1 flagellar protein of Salmonella typhimurium. I. The tryptic peptides. J Biol Chem. 1972 Aug 25;247(16):5180–5193. [PubMed] [Google Scholar]
  21. Kuwajima G. Construction of a minimum-size functional flagellin of Escherichia coli. J Bacteriol. 1988 Jul;170(7):3305–3309. doi: 10.1128/jb.170.7.3305-3309.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kuwajima G. Flagellin domain that affects H antigenicity of Escherichia coli K-12. J Bacteriol. 1988 Jan;170(1):485–488. doi: 10.1128/jb.170.1.485-488.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. Lee A., O'Rourke J. L., Barrington P. J., Trust T. J. Mucus colonization as a determinant of pathogenicity in intestinal infection by Campylobacter jejuni: a mouse cecal model. Infect Immun. 1986 Feb;51(2):536–546. doi: 10.1128/iai.51.2.536-546.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lior H., Woodward D. L., Edgar J. A., Laroche L. J., Gill P. Serotyping of Campylobacter jejuni by slide agglutination based on heat-labile antigenic factors. J Clin Microbiol. 1982 May;15(5):761–768. doi: 10.1128/jcm.15.5.761-768.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lipman D. J., Pearson W. R. Rapid and sensitive protein similarity searches. Science. 1985 Mar 22;227(4693):1435–1441. doi: 10.1126/science.2983426. [DOI] [PubMed] [Google Scholar]
  28. Logan S. M., Harris L. A., Trust T. J. Isolation and characterization of Campylobacter flagellins. J Bacteriol. 1987 Nov;169(11):5072–5077. doi: 10.1128/jb.169.11.5072-5077.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. 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]
  31. Logan S. M., Trust T. J. Outer membrane characteristics of Campylobacter jejuni. Infect Immun. 1982 Dec;38(3):898–906. doi: 10.1128/iai.38.3.898-906.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. McKern N. M., O'Donnell I. J., Inglis A. S., Stewart D. J., Clark B. L. Amino acid sequence of pilin from Bacteroides nodosus (strain 198), the causative organism of ovine footrot. FEBS Lett. 1983 Nov 28;164(1):149–153. doi: 10.1016/0014-5793(83)80039-8. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Nussenzweig V., Nussenzweig R. S. Circumsporozoite proteins of malaria parasites. Cell. 1985 Sep;42(2):401–403. doi: 10.1016/0092-8674(85)90093-5. [DOI] [PubMed] [Google Scholar]
  35. Owen R. J. Nucleic acids in the classification of Campylobacters. Eur J Clin Microbiol. 1983 Aug;2(4):367–377. doi: 10.1007/BF02019473. [DOI] [PubMed] [Google Scholar]
  36. Sastry P. A., Finlay B. B., Pasloske B. L., Paranchych W., Pearlstone J. R., Smillie L. B. Comparative studies of the amino acid and nucleotide sequences of pilin derived from Pseudomonas aeruginosa PAK and PAO. J Bacteriol. 1985 Nov;164(2):571–577. doi: 10.1128/jb.164.2.571-577.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Scott J. R., Hollingshead S. K., Fischetti V. A. Homologous regions within M protein genes in group A streptococci of different serotypes. Infect Immun. 1986 May;52(2):609–612. doi: 10.1128/iai.52.2.609-612.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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]
  39. Skirrow M. B. Campylobacter enteritis: a "new" disease. Br Med J. 1977 Jul 2;2(6078):9–11. doi: 10.1136/bmj.2.6078.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Taylor R. K., Miller V. L., Furlong D. B., Mekalanos J. J. Use of phoA gene fusions to identify a pilus colonization factor coordinately regulated with cholera toxin. Proc Natl Acad Sci U S A. 1987 May;84(9):2833–2837. doi: 10.1073/pnas.84.9.2833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Walker R. I., Caldwell M. B., Lee E. C., Guerry P., Trust T. J., Ruiz-Palacios G. M. Pathophysiology of Campylobacter enteritis. Microbiol Rev. 1986 Mar;50(1):81–94. doi: 10.1128/mr.50.1.81-94.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wei L. N., Joys T. M. Covalent structure of three phase-1 flagellar filament proteins of Salmonella. J Mol Biol. 1985 Dec 20;186(4):791–803. doi: 10.1016/0022-2836(85)90397-3. [DOI] [PubMed] [Google Scholar]
  43. Wenman W. M., Chai J., Louie T. J., Goudreau C., Lior H., Newell D. G., Pearson A. D., Taylor D. E. Antigenic analysis of Campylobacter flagellar protein and other proteins. J Clin Microbiol. 1985 Jan;21(1):108–112. doi: 10.1128/jcm.21.1.108-112.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]

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