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. 1987 Aug;169(8):3617–3624. doi: 10.1128/jb.169.8.3617-3624.1987

Identification of flagellar hook and basal body gene products (FlaFV, FlaFVI, FlaFVII and FlaFVIII) in Salmonella typhimurium.

M Homma, K Ohnishi, T Iino, R M Macnab
PMCID: PMC212441  PMID: 3301807

Abstract

The flagellar genes flaFV, flaFVII, and flaFVIII of Salmonella typhimurium were cloned, and their presence on a given plasmid was verified by complementation of Escherichia coli mutants defective in the homologous genes. The gene products were identified by radiolabeling in a minicell system as being proteins of the following molecular masses: FlaFV, 42 kilodaltons (kDa); FlaFVI, 32 kDa; FlaFVII, 30 kDA; and FlaFVIII, 27 kDa. These data, together with isoelectric focusing data, confirm gene product assignments of flagellar components made indirectly from mutant studies. Flagellar components are transported by either a signal peptide-dependent or a flagellar-specific pathway. Consistent with its location in the outer membrane ring of the basal body, protein FlaFVIII seems to use the signal peptide-dependent pathway, since it was synthesized in a precursor form and processed, presumably by peptide cleavage, to a mature form; the maturation process was inhibited by addition of a proton ionophore. Proteins synthesized in minicells were localized as follows: FlaFVI was localized to the soluble fraction (cytoplasm); pre-FlaFVIII and FlaFVIII were localized to the particulate fraction (membrane or high-molecular-weight aggregate); FlaFV and FlaFVII were localized to both fractions. The significance of these locations in terms of known or suspected roles in the flagellar apparatus is discussed.

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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. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  3. Daniels C. J., Bole D. G., Quay S. C., Oxender D. L. Role for membrane potential in the secretion of protein into the periplasm of Escherichia coli. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5396–5400. doi: 10.1073/pnas.78.9.5396. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DeFranco A. L., Parkinson J. S., Koshland D. E., Jr Functional homology of chemotaxis genes in Escherichia coli and Salmonella typhimurium. J Bacteriol. 1979 Jul;139(1):107–114. doi: 10.1128/jb.139.1.107-114.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. DePamphilis M. L., Adler J. Attachment of flagellar basal bodies to the cell envelope: specific attachment to the outer, lipopolysaccharide membrane and the cyoplasmic membrane. J Bacteriol. 1971 Jan;105(1):396–407. doi: 10.1128/jb.105.1.396-407.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Homma M., Fujita H., Yamaguchi S., Iino T. Excretion of unassembled flagellin by Salmonella typhimurium mutants deficient in hook-associated proteins. J Bacteriol. 1984 Sep;159(3):1056–1059. doi: 10.1128/jb.159.3.1056-1059.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Homma M., Iino T. Excretion of unassembled hook-associated proteins by Salmonella typhimurium. J Bacteriol. 1985 Dec;164(3):1370–1372. doi: 10.1128/jb.164.3.1370-1372.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Homma M., Iino T., Kutsukake K., Yamaguchi S. In vitro reconstitution of flagellar filaments onto hooks of filamentless mutants of Salmonella typhimurium by addition of hook-associated proteins. Proc Natl Acad Sci U S A. 1986 Aug;83(16):6169–6173. doi: 10.1073/pnas.83.16.6169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Homma M., Iino T. Locations of hook-associated proteins in flagellar structures of Salmonella typhimurium. J Bacteriol. 1985 Apr;162(1):183–189. doi: 10.1128/jb.162.1.183-189.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Homma M., Komeda Y., Iino T., Macnab R. M. The flaFIX gene product of Salmonella typhimurium is a flagellar basal body component with a signal peptide for export. J Bacteriol. 1987 Apr;169(4):1493–1498. doi: 10.1128/jb.169.4.1493-1498.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Homma M., Kutsukake K., Iino T. Structural genes for flagellar hook-associated proteins in Salmonella typhimurium. J Bacteriol. 1985 Aug;163(2):464–471. doi: 10.1128/jb.163.2.464-471.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Ikeda T., Homma M., Iino T., Asakura S., Kamiya R. Localization and stoichiometry of hook-associated proteins within Salmonella typhimurium flagella. J Bacteriol. 1987 Mar;169(3):1168–1173. doi: 10.1128/jb.169.3.1168-1173.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jones C. J., Homma M., Macnab R. M. Identification of proteins of the outer (L and P) rings of the flagellar basal body of Escherichia coli. J Bacteriol. 1987 Apr;169(4):1489–1492. doi: 10.1128/jb.169.4.1489-1492.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kagawa H., Owaribe K., Asakura S., Takahashi N. Flagellar hook protein from Salmonella SJ25. J Bacteriol. 1976 Jan;125(1):68–73. doi: 10.1128/jb.125.1.68-73.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Komeda Y. Fusions of flagellar operons to lactose genes on a mu lac bacteriophage. J Bacteriol. 1982 Apr;150(1):16–26. doi: 10.1128/jb.150.1.16-26.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Komeda Y., Silverman M., Matsumura P., Simon M. Genes for the hook-basal body proteins of the flagellar apparatus in Escherichia coli. J Bacteriol. 1978 May;134(2):655–667. doi: 10.1128/jb.134.2.655-667.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Komeda Y., Silverman M., Simon M. Identification of the structural gene for the hook subunit protein of Escherichia coli flagella. J Bacteriol. 1978 Jan;133(1):364–371. doi: 10.1128/jb.133.1.364-371.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Koshland D., Botstein D. Evidence for posttranslational translocation of beta-lactamase across the bacterial inner membrane. Cell. 1982 Oct;30(3):893–902. doi: 10.1016/0092-8674(82)90294-x. [DOI] [PubMed] [Google Scholar]
  21. Kutsukake K., Iino T. Refined genetic analysis of the region II che mutants in Salmonella typhimurium. Mol Gen Genet. 1985;199(3):406–409. doi: 10.1007/BF00330750. [DOI] [PubMed] [Google Scholar]
  22. Kutsukake K., Nakao T., Iino T. A gene for DNA invertase and an invertible DNA in Escherichia coli K-12. Gene. 1985;34(2-3):343–350. doi: 10.1016/0378-1119(85)90143-x. [DOI] [PubMed] [Google Scholar]
  23. Kutsukake K., Suzuki T., Yamaguchi S., Iino T. Role of gene flaFV on flagellar hook formation in Salmonella typhimurium. J Bacteriol. 1979 Oct;140(1):267–275. doi: 10.1128/jb.140.1.267-275.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Matsumura P., Silverman M., Simon M. Cloning and expression of the flagellar hook gene on hybird plasmids in minicells. Nature. 1977 Feb 24;265(5596):758–760. doi: 10.1038/265758a0. [DOI] [PubMed] [Google Scholar]
  25. Oliver D. Protein secretion in Escherichia coli. Annu Rev Microbiol. 1985;39:615–648. doi: 10.1146/annurev.mi.39.100185.003151. [DOI] [PubMed] [Google Scholar]
  26. Silverman M., Simon M. I. Bacterial flagella. Annu Rev Microbiol. 1977;31:397–419. doi: 10.1146/annurev.mi.31.100177.002145. [DOI] [PubMed] [Google Scholar]
  27. Smith R. W., Koffler H. Bacterial flagella. Adv Microb Physiol. 1971;6:219–339. doi: 10.1016/s0065-2911(08)60070-3. [DOI] [PubMed] [Google Scholar]
  28. Suzuki T., Iino T., Horiguchi T., Yamaguchi S. Incomplete flagellar structures in nonflagellate mutants of Salmonella typhimurium. J Bacteriol. 1978 Feb;133(2):904–915. doi: 10.1128/jb.133.2.904-915.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Yamaguchi S., Fujita H., Taira T., Kutsukake K., Homma M., Iino T. Genetic analysis of three additional fla genes in Salmonella typhimurium. J Gen Microbiol. 1984 Dec;130(12):3339–3342. doi: 10.1099/00221287-130-12-3339. [DOI] [PubMed] [Google Scholar]

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