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J Bacteriol. Jun 1995; 177(11): 3010–3020.
PMCID: PMC176987

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.


Helicobacter mustelae causes chronic gastritis and ulcer disease in ferrets. It is therefore considered an important animal model of human Helicobacter pylori infection. High motility even in a viscous environment is one of the common virulence determinants of Helicobacter species. Their sheathed flagella contain a complex filament that is composed of two distinctly different flagellin subunits, FlaA and FlaB, that are coexpressed in different amounts. Here, we report the cloning and sequence determination of the flaA gene of H. mustelae NCTC12032 from a PCR amplification product. The FlaA protein has a calculated molecular mass of 53 kDa and is 73% homologous to the H. pylori FlaA subunit. Isogenic flaA and flaB mutants of H. mustelae F1 were constructed by means of reverse genetics. A method was established to generate double mutants (flaA flaB) of H. mustelae F1 as well as H. pylori N6. Genotypes, motility properties, and morphologies of the H. mustelae flagellin mutants were determined and compared with those of the H. pylori flaA and flaB mutants described previously. The flagellar organizations of the two Helicobacter species proved to be highly similar. When the flaB genes were disrupted, motility decreased by 30 to 40%. flaA mutants retained weak motility by comparison with strains that were devoid of both flagellin subunits. Weakly positive motility tests of the flaA mutants correlated with the existence of short truncated flagella. In H. mustelae, lateral as well as polar flagella were present in the truncated form. flaA flaB double mutants were completely nonmotile and lacked any form of flagella. These results show that the presence of both flagellin subunits is necessary for complete motility of Helicobacter species. The importance of this flagellar organization for the ability of the bacteria to colonize the gastric mucosa and to persist in the gastric mucus remains to be proven.

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Selected References

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  • Alm RA, Guerry P, Trust TJ. The Campylobacter sigma 54 flaB flagellin promoter is subject to environmental regulation. J Bacteriol. 1993 Jul;175(14):4448–4455. [PMC free article] [PubMed]
  • Alm RA, Guerry P, Trust TJ. Significance of duplicated flagellin genes in Campylobacter. J Mol Biol. 1993 Mar 20;230(2):359–363. [PubMed]
  • Blaser MJ. Helicobacter pylori: microbiology of a 'slow' bacterial infection. Trends Microbiol. 1993 Oct;1(7):255–260. [PubMed]
  • Boyer HW, Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. [PubMed]
  • Casadaban MJ, Cohen SN. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol. 1980 Apr;138(2):179–207. [PubMed]
  • Cover TL, Blaser MJ. Helicobacter pylori and gastroduodenal disease. Annu Rev Med. 1992;43:135–145. [PubMed]
  • Driks A, Bryan R, Shapiro L, DeRosier DJ. The organization of the Caulobacter crescentus flagellar filament. J Mol Biol. 1989 Apr 20;206(4):627–636. [PubMed]
  • Dunn BE, Sung CC, Taylor NS, Fox JG. Purification and characterization of Helicobacter mustelae urease. Infect Immun. 1991 Sep;59(9):3343–3345. [PMC free article] [PubMed]
  • Eaton KA, Brooks CL, Morgan DR, Krakowka S. Essential role of urease in pathogenesis of gastritis induced by Helicobacter pylori in gnotobiotic piglets. Infect Immun. 1991 Jul;59(7):2470–2475. [PMC free article] [PubMed]
  • Eaton KA, Morgan DR, Krakowka S. Campylobacter pylori virulence factors in gnotobiotic piglets. Infect Immun. 1989 Apr;57(4):1119–1125. [PMC free article] [PubMed]
  • Eaton KA, Morgan DR, Krakowka S. Motility as a factor in the colonisation of gnotobiotic piglets by Helicobacter pylori. J Med Microbiol. 1992 Aug;37(2):123–127. [PubMed]
  • Evans DG, Evans DJ, Jr, Moulds JJ, Graham DY. N-acetylneuraminyllactose-binding fibrillar hemagglutinin of Campylobacter pylori: a putative colonization factor antigen. Infect Immun. 1988 Nov;56(11):2896–2906. [PMC free article] [PubMed]
  • Ferrero RL, Cussac V, Courcoux P, Labigne A. Construction of isogenic urease-negative mutants of Helicobacter pylori by allelic exchange. J Bacteriol. 1992 Jul;174(13):4212–4217. [PMC free article] [PubMed]
  • Ferrero RL, 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. [PubMed]
  • Fox JG, Cabot EB, Taylor NS, Laraway R. Gastric colonization by Campylobacter pylori subsp. mustelae in ferrets. Infect Immun. 1988 Nov;56(11):2994–2996. [PMC free article] [PubMed]
  • Fox JG, Correa P, Taylor NS, Lee A, Otto G, Murphy JC, Rose R. Helicobacter mustelae-associated gastritis in ferrets. An animal model of Helicobacter pylori gastritis in humans. Gastroenterology. 1990 Aug;99(2):352–361. [PubMed]
  • Fox JG, Otto G, Murphy JC, Taylor NS, Lee A. Gastric colonization of the ferret with Helicobacter species: natural and experimental infections. Rev Infect Dis. 1991 Jul-Aug;13 (Suppl 8):S671–S680. [PubMed]
  • Geis G, Leying H, Suerbaum S, Mai U, Opferkuch W. Ultrastructure and chemical analysis of Campylobacter pylori flagella. J Clin Microbiol. 1989 Mar;27(3):436–441. [PMC free article] [PubMed]
  • Geis G, Suerbaum S, Forsthoff B, Leying H, Opferkuch W. Ultrastructure and biochemical studies of the flagellar sheath of Helicobacter pylori. J Med Microbiol. 1993 May;38(5):371–377. [PubMed]
  • Gerl L, Sumper M. Halobacterial flagellins are encoded by a multigene family. Characterization of five flagellin genes. J Biol Chem. 1988 Sep 15;263(26):13246–13251. [PubMed]
  • Goodwin CS, McCulloch RK, Armstrong JA, Wee SH. Unusual cellular fatty acids and distinctive ultrastructure in a new spiral bacterium (Campylobacter pyloridis) from the human gastric mucosa. J Med Microbiol. 1985 Apr;19(2):257–267. [PubMed]
  • Guerry P, Alm RA, Power ME, Logan SM, Trust TJ. Role of two flagellin genes in Campylobacter motility. J Bacteriol. 1991 Aug;173(15):4757–4764. [PMC free article] [PubMed]
  • Guerry P, Logan SM, Thornton S, Trust TJ. Genomic organization and expression of Campylobacter flagellin genes. J Bacteriol. 1990 Apr;172(4):1853–1860. [PMC free article] [PubMed]
  • Haas R, Meyer TF, van Putten JP. Aflagellated mutants of Helicobacter pylori generated by genetic transformation of naturally competent strains using transposon shuttle mutagenesis. Mol Microbiol. 1993 May;8(4):753–760. [PubMed]
  • Hazell SL, Lee A, Brady L, Hennessy W. Campylobacter pyloridis and gastritis: association with intercellular spaces and adaptation to an environment of mucus as important factors in colonization of the gastric epithelium. J Infect Dis. 1986 Apr;153(4):658–663. [PubMed]
  • Kostrzynska M, Betts JD, Austin JW, Trust TJ. Identification, characterization, and spatial localization of two flagellin species in Helicobacter pylori flagella. J Bacteriol. 1991 Feb;173(3):937–946. [PMC free article] [PubMed]
  • Krakowka S, Morgan DR, Kraft WG, Leunk RD. Establishment of gastric Campylobacter pylori infection in the neonatal gnotobiotic piglet. Infect Immun. 1987 Nov;55(11):2789–2796. [PMC free article] [PubMed]
  • Labigne A, Cussac V, Courcoux P. Shuttle cloning and nucleotide sequences of Helicobacter pylori genes responsible for urease activity. J Bacteriol. 1991 Mar;173(6):1920–1931. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Lee A, Fox JG, Otto G, Murphy J. A small animal model of human Helicobacter pylori active chronic gastritis. Gastroenterology. 1990 Nov;99(5):1315–1323. [PubMed]
  • Leying H, Suerbaum S, Geis G, Haas R. Cloning and genetic characterization of a Helicobacter pylori flagellin gene. Mol Microbiol. 1992 Oct;6(19):2863–2874. [PubMed]
  • Lugtenberg B, Meijers J, Peters R, van der Hoek P, van Alphen L. Electrophoretic resolution of the "major outer membrane protein" of Escherichia coli K12 into four bands. FEBS Lett. 1975 Oct 15;58(1):254–258. [PubMed]
  • Majewski SI, Goodwin CS. Restriction endonuclease analysis of the genome of Campylobacter pylori with a rapid extraction method: evidence for considerable genomic variation. J Infect Dis. 1988 Mar;157(3):465–471. [PubMed]
  • Messing J, Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. [PubMed]
  • Minnich SA, Ohta N, Taylor N, Newton A. Role of the 25-, 27-, and 29-kilodalton flagellins in Caulobacter crescentus cell motility: method for construction of deletion and Tn5 insertion mutants by gene replacement. J Bacteriol. 1988 Sep;170(9):3953–3960. [PMC free article] [PubMed]
  • Nuijten PJ, van Asten FJ, Gaastra W, van der Zeijst BA. Structural and functional analysis of two Campylobacter jejuni flagellin genes. J Biol Chem. 1990 Oct 15;265(29):17798–17804. [PubMed]
  • O'Rourke J, Lee A, Fox JG. An ultrastructural study of Helicobacter mustelae and evidence of a specific association with gastric mucosa. J Med Microbiol. 1992 Jun;36(6):420–427. [PubMed]
  • Parsonnet J. Helicobacter pylori and gastric cancer. Gastroenterol Clin North Am. 1993 Mar;22(1):89–104. [PubMed]
  • Pleier E, Schmitt R. Identification and sequence analysis of two related flagellin genes in Rhizobium meliloti. J Bacteriol. 1989 Mar;171(3):1467–1475. [PMC free article] [PubMed]
  • Pleier E, Schmitt R. Expression of two Rhizobium meliloti flagellin genes and their contribution to the complex filament structure. J Bacteriol. 1991 Mar;173(6):2077–2085. [PMC free article] [PubMed]
  • Richardson K, Nixon L, Mostow P, Kaper JB, Michalski J. Transposon-induced non-motile mutants of Vibrio cholerae. J Gen Microbiol. 1990 Apr;136(4):717–725. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. [PubMed]
  • Suerbaum S, Geis G, Josenhans C, Opferkuch W. Biochemical studies of Helicobacter mustelae fatty acid composition and flagella. Infect Immun. 1992 Apr;60(4):1695–1698. [PMC free article] [PubMed]
  • Suerbaum S, Josenhans C, Labigne A. Cloning and genetic characterization of the Helicobacter pylori and Helicobacter mustelae flaB flagellin genes and construction of H. pylori flaA- and flaB-negative mutants by electroporation-mediated allelic exchange. J Bacteriol. 1993 Jun;175(11):3278–3288. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Trachtenberg S, DeRosier DJ, Macnab RM. Three-dimensional structure of the complex flagellar filament of Rhizobium lupini and its relation to the structure of the plain filament. J Mol Biol. 1987 Jun 5;195(3):603–620. [PubMed]
  • Trieu-Cuot P, Gerbaud G, Lambert T, Courvalin P. In vivo transfer of genetic information between gram-positive and gram-negative bacteria. EMBO J. 1985 Dec 16;4(13A):3583–3587. [PMC free article] [PubMed]
  • Turbett GR, Høj PB, Horne R, Mee BJ. Purification and characterization of the urease enzymes of Helicobacter species from humans and animals. Infect Immun. 1992 Dec;60(12):5259–5266. [PMC free article] [PubMed]
  • Wilson DR, Beveridge TJ. Bacterial flagellar filaments and their component flagellins. Can J Microbiol. 1993 May;39(5):451–472. [PubMed]
  • 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. [PubMed]

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