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. 1992 Oct;174(19):6046–6053. doi: 10.1128/jb.174.19.6046-6053.1992

The Caulobacter crescentus flaFG region regulates synthesis and assembly of flagellin proteins encoded by two genetically unlinked gene clusters.

P V Schoenlein 1, J Lui 1, L Gallman 1, B Ely 1
PMCID: PMC207669  PMID: 1400155

Abstract

At a specific time in the Caulobacter crescentus cell cycle, a single flagellar filament and multiple receptor sites for the swarmer-specific phage phi Cbk are assembled at one pole of the predivisional cell. One cluster of genes required for this morphogenesis, the flaYG region, includes the flgJKL genes, which encode structural proteins of the flagellar filament. These flagellin genes are flanked by genes required for filament assembly, the flaYE genes at one end and the flaF-flbT-flbA-flaG genes at the other. In this study, we characterized mutants carrying large chromosomal deletions within this region. Several of these strains are phi CbK resistant and produce a novel 22-kDa flagellin that is not assembled into flagella. Merodiploid strains containing either the entire flaFG region or individual fla transcription units from this region were constructed. These strains were used to correlate the presence or absence of specific gene products to changes in flagellin synthesis, filament assembly, or phage sensitivity. As a result of these studies, we were able to conclude that (i) the production of the 22-kDa flagellin results from the absence of the flbA and flaG gene products, which appear to be components of a flagellin-processing pathway common to the 25-, 27-, and 29-kDa flagellins; (ii) flbT negatively modulates the synthesis of the 27- and 25-kDa flagellins from two genetically unlinked gene clusters; (iii) flgL is the only flagellin gene able to encode the 27-kDa flagellin, and this flagellin appears to be required for the efficient assembly of the 25-kDa flagellins; (iv) flaF is required for filament assembly; and (v) phi CbK resistance results from the deletion of at least two genes in the flaFG region.

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Selected References

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  1. Barrett J. T., Croft R. H., Ferber D. M., Gerardot C. J., Schoenlein P. V., Ely B. Genetic mapping with Tn5-derived auxotrophs of Caulobacter crescentus. J Bacteriol. 1982 Aug;151(2):888–898. doi: 10.1128/jb.151.2.888-898.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bender R. A., Refson C. M., O'Neill E. A. Role of the flagellum in cell-cycle-dependent expression of bacteriophage receptor activity in Caulobacter crescentus. J Bacteriol. 1989 Feb;171(2):1035–1040. doi: 10.1128/jb.171.2.1035-1040.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Driks A., Bryan R., Shapiro L., DeRosier D. J. The organization of the Caulobacter crescentus flagellar filament. J Mol Biol. 1989 Apr 20;206(4):627–636. doi: 10.1016/0022-2836(89)90571-8. [DOI] [PubMed] [Google Scholar]
  4. Ely B., Croft R. H., Gerardot C. J. Genetic mapping of genes required for motility in Caulobacter crescentus. Genetics. 1984 Nov;108(3):523–532. doi: 10.1093/genetics/108.3.523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ely B., Croft R. H. Transposon mutagenesis in Caulobacter crescentus. J Bacteriol. 1982 Feb;149(2):620–625. doi: 10.1128/jb.149.2.620-625.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ely B., Ely T. W. Use of pulsed field gel electrophoresis and transposon mutagenesis to estimate the minimal number of genes required for motility in Caulobacter crescentus. Genetics. 1989 Dec;123(4):649–654. doi: 10.1093/genetics/123.4.649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ely B., Gerardot C. J. Use of pulsed-field-gradient gel electrophoresis to construct a physical map of the Caulobacter crescentus genome. Gene. 1988 Sep 7;68(2):323–333. doi: 10.1016/0378-1119(88)90035-2. [DOI] [PubMed] [Google Scholar]
  8. Ely B., Johnson R. C. Generalized Transduction in CAULOBACTER CRESCENTUS. Genetics. 1977 Nov;87(3):391–399. doi: 10.1093/genetics/87.3.391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fukuda A., Asada M., Koyasu S., Yoshida H., Yaginuma K., Okada Y. Regulation of polar morphogenesis in Caulobacter crescentus. J Bacteriol. 1981 Jan;145(1):559–572. doi: 10.1128/jb.145.1.559-572.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gill P. R., Agabian N. The nucleotide sequence of the Mr = 28,500 flagellin gene of Caulobacter crescentus. J Biol Chem. 1983 Jun 25;258(12):7395–7401. [PubMed] [Google Scholar]
  11. Hahnenberger K. M., Shapiro L. Identification of a gene cluster involved in flagellar basal body biogenesis in Caulobacter crescentus. J Mol Biol. 1987 Mar 5;194(1):91–103. doi: 10.1016/0022-2836(87)90718-2. [DOI] [PubMed] [Google Scholar]
  12. Heimbrook M. E., Wang W. L., Campbell G. Staining bacterial flagella easily. J Clin Microbiol. 1989 Nov;27(11):2612–2615. doi: 10.1128/jcm.27.11.2612-2615.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Johnson R. C., Ely B. Analysis of nonmotile mutants of the dimorphic bacterium Caulobacter crescentus. J Bacteriol. 1979 Jan;137(1):627–634. doi: 10.1128/jb.137.1.627-634.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Johnson R. C., Ferber D. M., Ely B. Synthesis and assembly of flagellar components by Caulobacter crescentus motility mutants. J Bacteriol. 1983 Jun;154(3):1137–1144. doi: 10.1128/jb.154.3.1137-1144.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Johnson R. C., Walsh M. P., Ely B., Shapiro L. Flagellar hook and basal complex of Caulobacter crescentus. J Bacteriol. 1979 Jun;138(3):984–989. doi: 10.1128/jb.138.3.984-989.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Khambaty F. M., Ely B. Molecular genetics of the flgI region and its role in flagellum biosynthesis in Caulobacter crescentus. J Bacteriol. 1992 Jun;174(12):4101–4109. doi: 10.1128/jb.174.12.4101-4109.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Koyasu S., Asada M., Fukuda A., Okada Y. Sequential polymerization of flagellin A and flagellin B into Caulobacter flagella. J Mol Biol. 1981 Dec 5;153(2):471–475. doi: 10.1016/0022-2836(81)90292-8. [DOI] [PubMed] [Google Scholar]
  18. Koyasu S. On flagellar formation in Caulobacter crescentus: novel flagellin synthesis in stub-forming non-motile mutants of C. crescentus. J Biochem. 1984 Nov;96(5):1351–1364. doi: 10.1093/oxfordjournals.jbchem.a134963. [DOI] [PubMed] [Google Scholar]
  19. Minnich S. A., Newton A. Promoter mapping and cell cycle regulation of flagellin gene transcription in Caulobacter crescentus. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1142–1146. doi: 10.1073/pnas.84.5.1142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ohta N., Swanson E., Ely B., Newton A. Physical mapping and complementation analysis of transposon Tn5 mutations in Caulobacter crescentus: organization of transcriptional units in the hook gene cluster. J Bacteriol. 1984 Jun;158(3):897–904. doi: 10.1128/jb.158.3.897-904.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. POINDEXTER J. S. BIOLOGICAL PROPERTIES AND CLASSIFICATION OF THE CAULOBACTER GROUP. Bacteriol Rev. 1964 Sep;28:231–295. doi: 10.1128/br.28.3.231-295.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Purucker M., Bryan R., Amemiya K., Ely B., Shapiro L. Isolation of a Caulobacter gene cluster specifying flagellum production by using nonmotile Tn5 insertion mutants. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6797–6801. doi: 10.1073/pnas.79.22.6797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schoenlein P. V., Ely B. Characterization of strains containing mutations in the contiguous flaF, flbT, or flbA-flaG transcription unit and identification of a novel fla phenotype in Caulobacter crescentus. J Bacteriol. 1989 Mar;171(3):1554–1561. doi: 10.1128/jb.171.3.1554-1561.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schoenlein P. V., Gallman L. S., Ely B. Organization of the flaFG gene cluster and identification of two additional genes involved in flagellum biogenesis in Caulobacter crescentus. J Bacteriol. 1989 Mar;171(3):1544–1553. doi: 10.1128/jb.171.3.1544-1553.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schoenlein P. V., Gallman L. S., Winkler M. E., Ely B. Nucleotide sequence of the Caulobacter crescentus flaF and flbT genes and an analysis of codon usage in organisms with G + C-rich genomes. Gene. 1990 Sep 1;93(1):17–25. doi: 10.1016/0378-1119(90)90130-j. [DOI] [PubMed] [Google Scholar]
  26. Smith M. R., Devine C. S., Cohn S. M., Lieberman M. W. Quantitative electrophoretic transfer of DNA from polyacrylamide or agarose gels to nitrocellulose. Anal Biochem. 1984 Feb;137(1):120–124. doi: 10.1016/0003-2697(84)90356-7. [DOI] [PubMed] [Google Scholar]
  27. Tarleton J. C., Ely B. Isolation and characterization of ilvA, ilvBN, and ilvD mutants of Caulobacter crescentus. J Bacteriol. 1991 Feb;173(3):1259–1267. doi: 10.1128/jb.173.3.1259-1267.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
  29. 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]
  30. Winkler M. E., Schoenlein P. V., Ross C. M., Barrett J. T., Ely B. Genetic and physical analyses of Caulobacter crescentus trp genes. J Bacteriol. 1984 Oct;160(1):279–287. doi: 10.1128/jb.160.1.279-287.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]

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