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. 1997 Sep;179(17):5292–5299. doi: 10.1128/jb.179.17.5292-5299.1997

Identification and molecular characterization of two tandemly located flagellin genes from Aeromonas salmonicida A449.

E Umelo 1, T J Trust 1
PMCID: PMC179395  PMID: 9286979

Abstract

Two tandemly located flagellin genes, flaA and flaB, with 79% nucleotide sequence identity were identified in Aeromonas salmonicida A449. The fla genes are conserved in typical and atypical strains of A. salmonicida, and they display significant divergence at the nucleotide level from the fla genes of the motile species Aeromonas hydrophila and Aeromonas veronii biotype sobria. flaA and flaB encode unprocessed flagellins with predicted Mrs of 32,351 and 32,056, respectively. When cloned under the control of the Ptac promoter, flaB was highly expressed when induced in Escherichia coli DH5alpha, and the FlaB protein was detectable even in the uninduced state. In flaA clones containing intact upstream sequence, FlaA was barely detectable when uninduced and poorly expressed on induction. The A. salmonicida flagellins are antigenically cross-reactive with the A. hydrophila TF7 flagellin(s) and evolutionarily closely related to the flagellins of Pseudomonas aeruginosa and Vibrio anguillarum. Electron microscopy showed that A. salmonicida A449 expresses unsheathed polar flagella at an extremely low frequency under normal laboratory growth conditions, suggesting the presence of a full complement of genes whose products are required to make flagella; e.g., immediately downstream of flaA and flaB are open reading frames encoding FlaG and FlaH homologs.

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Selected References

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  1. Akerley B. J., Monack D. M., Falkow S., Miller J. F. The bvgAS locus negatively controls motility and synthesis of flagella in Bordetella bronchiseptica. J Bacteriol. 1992 Feb;174(3):980–990. doi: 10.1128/jb.174.3.980-990.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Al Mamun A. A., Tominaga A., Enomoto M. Detection and characterization of the flagellar master operon in the four Shigella subgroups. J Bacteriol. 1996 Jul;178(13):3722–3726. doi: 10.1128/jb.178.13.3722-3726.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  4. Attridge S. R., Rowley D. The role of the flagellum in the adherence of Vibrio cholerae. J Infect Dis. 1983 May;147(5):864–872. doi: 10.1093/infdis/147.5.864. [DOI] [PubMed] [Google Scholar]
  5. Belland R. J., Trust T. J. DNA:DNA reassociation analysis of Aeromonas salmonicida. J Gen Microbiol. 1988 Feb;134(2):307–315. doi: 10.1099/00221287-134-2-307. [DOI] [PubMed] [Google Scholar]
  6. Bryant T. N., Lee J. V., West P. A., Colwell R. R. Numerical classification of species of Vibrio and related genera. J Appl Bacteriol. 1986 Nov;61(5):437–467. doi: 10.1111/j.1365-2672.1986.tb04308.x. [DOI] [PubMed] [Google Scholar]
  7. Drake D., Montie T. C. Flagella, motility and invasive virulence of Pseudomonas aeruginosa. J Gen Microbiol. 1988 Jan;134(1):43–52. doi: 10.1099/00221287-134-1-43. [DOI] [PubMed] [Google Scholar]
  8. Eaton K. A., Morgan D. R., Krakowka S. Motility as a factor in the colonisation of gnotobiotic piglets by Helicobacter pylori. J Med Microbiol. 1992 Aug;37(2):123–127. doi: 10.1099/00222615-37-2-123. [DOI] [PubMed] [Google Scholar]
  9. Fürste J. P., Pansegrau W., Frank R., Blöcker H., Scholz P., Bagdasarian M., Lanka E. Molecular cloning of the plasmid RP4 primase region in a multi-host-range tacP expression vector. Gene. 1986;48(1):119–131. doi: 10.1016/0378-1119(86)90358-6. [DOI] [PubMed] [Google Scholar]
  10. Girón J. A. Expression of flagella and motility by Shigella. Mol Microbiol. 1995 Oct;18(1):63–75. doi: 10.1111/j.1365-2958.1995.mmi_18010063.x. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Gustafson C. E., Chu S., Trust T. J. Mutagenesis of the paracrystalline surface protein array of Aeromonas salmonicida by endogenous insertion elements. J Mol Biol. 1994 Apr 8;237(4):452–463. doi: 10.1006/jmbi.1994.1247. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. 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]
  16. Ishiguro E. E., Ainsworth T., Kay W. W., Trust T. J. Heme requirement for growth of fastidious atypical strains of Aeromonas salmonicida. Appl Environ Microbiol. 1986 Mar;51(3):668–670. doi: 10.1128/aem.51.3.668-670.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kostrzynska M., Betts J. D., Austin J. W., Trust T. J. Identification, characterization, and spatial localization of two flagellin species in Helicobacter pylori flagella. J Bacteriol. 1991 Feb;173(3):937–946. doi: 10.1128/jb.173.3.937-946.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Leigh A. F., Coote J. G., Parton R., Duggleby C. J. Chromosomal DNA from both flagellate and non-flagellate Bordetella species contains sequences homologous to the Salmonella H1 flagellin gene. FEMS Microbiol Lett. 1993 Aug 1;111(2-3):225–231. doi: 10.1111/j.1574-6968.1993.tb06390.x. [DOI] [PubMed] [Google Scholar]
  19. Li J., Nelson K., McWhorter A. C., Whittam T. S., Selander R. K. Recombinational basis of serovar diversity in Salmonella enterica. Proc Natl Acad Sci U S A. 1994 Mar 29;91(7):2552–2556. doi: 10.1073/pnas.91.7.2552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Martínez de Tejada G., Miller J. F., Cotter P. A. Comparative analysis of the virulence control systems of Bordetella pertussis and Bordetella bronchiseptica. Mol Microbiol. 1996 Dec;22(5):895–908. doi: 10.1046/j.1365-2958.1996.01538.x. [DOI] [PubMed] [Google Scholar]
  22. McCarter L. L. Genetic and molecular characterization of the polar flagellum of Vibrio parahaemolyticus. J Bacteriol. 1995 Mar;177(6):1595–1609. doi: 10.1128/jb.177.6.1595-1609.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. McGee K., Hörstedt P., Milton D. L. Identification and characterization of additional flagellin genes from Vibrio anguillarum. J Bacteriol. 1996 Sep;178(17):5188–5198. doi: 10.1128/jb.178.17.5188-5198.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. McIntosh D., Austin B. Atypical characteristics of the salmonid pathogen Aeromonas salmonicida. J Gen Microbiol. 1991 Jun;137(6):1341–1343. doi: 10.1099/00221287-137-6-1341. [DOI] [PubMed] [Google Scholar]
  25. Milton D. L., O'Toole R., Horstedt P., Wolf-Watz H. Flagellin A is essential for the virulence of Vibrio anguillarum. J Bacteriol. 1996 Mar;178(5):1310–1319. doi: 10.1128/jb.178.5.1310-1319.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Namba K., Yamashita I., Vonderviszt F. Structure of the core and central channel of bacterial flagella. Nature. 1989 Dec 7;342(6250):648–654. doi: 10.1038/342648a0. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Pleier E., Schmitt R. Identification and sequence analysis of two related flagellin genes in Rhizobium meliloti. J Bacteriol. 1989 Mar;171(3):1467–1475. doi: 10.1128/jb.171.3.1467-1475.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schoenhals G., Whitfield C. Comparative analysis of flagellin sequences from Escherichia coli strains possessing serologically distinct flagellar filaments with a shared complex surface pattern. J Bacteriol. 1993 Sep;175(17):5395–5402. doi: 10.1128/jb.175.17.5395-5402.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tominaga A., Mahmoud M. A., Mukaihara T., Enomoto M. Molecular characterization of intact, but cryptic, flagellin genes in the genus Shigella. Mol Microbiol. 1994 Apr;12(2):277–285. doi: 10.1111/j.1365-2958.1994.tb01016.x. [DOI] [PubMed] [Google Scholar]
  31. Totten P. A., Lory S. Characterization of the type a flagellin gene from Pseudomonas aeruginosa PAK. J Bacteriol. 1990 Dec;172(12):7188–7199. doi: 10.1128/jb.172.12.7188-7199.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Trust T. J., Klotz F. W., Miller L. H. Pathogenesis of infectious diseases of fish. Annu Rev Microbiol. 1986;40:479–502. doi: 10.1146/annurev.mi.40.100186.002403. [DOI] [PubMed] [Google Scholar]
  33. Umelo E., Noonan B., Trust T. J. Cloning, characterization and expression of the recA gene of Aeromonas salmonicida. Gene. 1996 Oct 10;175(1-2):133–136. doi: 10.1016/0378-1119(96)00138-2. [DOI] [PubMed] [Google Scholar]
  34. Weissborn A., Steinmann H. M., Shapiro L. Characterization of the proteins of the Caulobacter crescentus flagellar filament. Peptide analysis and filament organization. J Biol Chem. 1982 Feb 25;257(4):2066–2074. [PubMed] [Google Scholar]
  35. 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]
  36. 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]

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