Abstract
The fimA gene of Xanthomonas campestris pv. vesicatoria was identified and characterized. A 20-mer degenerate oligonucleotide complementary to the N-terminal amino acid sequence of the purified 15.5-kDa fimbrillin was used to locate fimA on a 2.6-kb SalI fragment of the X. campestris pv. vesicatoria 3240 genome. The nucleotide sequence of a 1.4-kb fragment containing the fimA region revealed two open reading frames predicting highly homologous proteins FimA and FimB. FimA, which was composed of 136 amino acids and had a calculated molecular weight of 14,302, showed high sequence identity to the type IV fimbrillin precursors. fimB predicted a protein product of 135 amino acids and a molecular weight of 13,854. The open reading frame for fimB contained near the 5' end a palindromic sequence with a terminator loop potential, and the expression level of fimB in vitro and in Xanthomonas was considerably lower than that of fimA. We detected an efficiently transcribed fimA-specific mRNA of 600 bases as well as two weakly expressed, longer mRNA species that reacted with both fimA and fimB. A homolog of fimA but not of fimB was detected by Southern hybridization in strains of X. campestris pv. vesicatoria, campestris, begoniae, translucens, and graminis. A fimA::omega mutant of strain 3240 was not significantly reduced in virulence or adhesiveness to tomato leaves. However, the fimA mutant was dramatically reduced in cell aggregation in laboratory cultures and on infected tomato leaves. The fimA mutant strain also exhibited decreased tolerance to UV light.
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- 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]
- Ausubel F. M. Regulation of nitrogen fixation genes. Cell. 1984 May;37(1):5–6. doi: 10.1016/0092-8674(84)90294-0. [DOI] [PubMed] [Google Scholar]
- Beattie G. A., Lindow S. E. Epiphytic fitness of phytopathogenic bacteria: physiological adaptations for growth and survival. Curr Top Microbiol Immunol. 1994;192:1–27. doi: 10.1007/978-3-642-78624-2_1. [DOI] [PubMed] [Google Scholar]
- Bonas U. hrp genes of phytopathogenic bacteria. Curr Top Microbiol Immunol. 1994;192:79–98. doi: 10.1007/978-3-642-78624-2_4. [DOI] [PubMed] [Google Scholar]
- Chen H. Z., Zubay G. Prokaryotic coupled transcription-translation. Methods Enzymol. 1983;101:674–690. doi: 10.1016/0076-6879(83)01047-2. [DOI] [PubMed] [Google Scholar]
- Chiang S. L., Taylor R. K., Koomey M., Mekalanos J. J. Single amino acid substitutions in the N-terminus of Vibrio cholerae TcpA affect colonization, autoagglutination, and serum resistance. Mol Microbiol. 1995 Sep;17(6):1133–1142. doi: 10.1111/j.1365-2958.1995.mmi_17061133.x. [DOI] [PubMed] [Google Scholar]
- Daniels M. J., Barber C. E., Turner P. C., Sawczyc M. K., Byrde R. J., Fielding A. H. Cloning of genes involved in pathogenicity of Xanthomonas campestris pv. campestris using the broad host range cosmid pLAFR1. EMBO J. 1984 Dec 20;3(13):3323–3328. doi: 10.1002/j.1460-2075.1984.tb02298.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Donnenberg M. S., Girón J. A., Nataro J. P., Kaper J. B. A plasmid-encoded type IV fimbrial gene of enteropathogenic Escherichia coli associated with localized adherence. Mol Microbiol. 1992 Nov;6(22):3427–3437. doi: 10.1111/j.1365-2958.1992.tb02210.x. [DOI] [PubMed] [Google Scholar]
- Dow J. M., Daniels M. J. Pathogenicity determinants and global regulation of pathogenicity of Xanthomonas campestris pv. campestris. Curr Top Microbiol Immunol. 1994;192:29–41. doi: 10.1007/978-3-642-78624-2_2. [DOI] [PubMed] [Google Scholar]
- Dums F., Dow J. M., Daniels M. J. Structural characterization of protein secretion genes of the bacterial phytopathogen Xanthomonas campestris pathovar campestris: relatedness to secretion systems of other gram-negative bacteria. Mol Gen Genet. 1991 Oct;229(3):357–364. doi: 10.1007/BF00267456. [DOI] [PubMed] [Google Scholar]
- Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
- Figurski D. H., Helinski D. R. Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1648–1652. doi: 10.1073/pnas.76.4.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Girón J. A., Ho A. S., Schoolnik G. K. An inducible bundle-forming pilus of enteropathogenic Escherichia coli. Science. 1991 Nov 1;254(5032):710–713. doi: 10.1126/science.1683004. [DOI] [PubMed] [Google Scholar]
- Higgins D. G., Sharp P. M. CLUSTAL: a package for performing multiple sequence alignment on a microcomputer. Gene. 1988 Dec 15;73(1):237–244. doi: 10.1016/0378-1119(88)90330-7. [DOI] [PubMed] [Google Scholar]
- Hu N. T., Hung M. N., Chiou S. J., Tang F., Chiang D. C., Huang H. Y., Wu C. Y. Cloning and characterization of a gene required for the secretion of extracellular enzymes across the outer membrane by Xanthomonas campestris pv. campestris. J Bacteriol. 1992 Apr;174(8):2679–2687. doi: 10.1128/jb.174.8.2679-2687.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knoop V., Staskawicz B., Bonas U. Expression of the avirulence gene avrBs3 from Xanthomonas campestris pv. vesicatoria is not under the control of hrp genes and is independent of plant factors. J Bacteriol. 1991 Nov;173(22):7142–7150. doi: 10.1128/jb.173.22.7142-7150.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Korhonen T. K., Nurmiaho E. L., Ranta H., Edén C. S. New Method for isolation of immunologically pure pili from Escherichia coli. Infect Immun. 1980 Feb;27(2):569–575. doi: 10.1128/iai.27.2.569-575.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Marceau M., Beretti J. L., Nassif X. High adhesiveness of encapsulated Neisseria meningitidis to epithelial cells is associated with the formation of bundles of pili. Mol Microbiol. 1995 Sep;17(5):855–863. doi: 10.1111/j.1365-2958.1995.mmi_17050855.x. [DOI] [PubMed] [Google Scholar]
- Marrs C. F., Schoolnik G., Koomey J. M., Hardy J., Rothbard J., Falkow S. Cloning and sequencing of a Moraxella bovis pilin gene. J Bacteriol. 1985 Jul;163(1):132–139. doi: 10.1128/jb.163.1.132-139.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nunn D., Bergman S., Lory S. Products of three accessory genes, pilB, pilC, and pilD, are required for biogenesis of Pseudomonas aeruginosa pili. J Bacteriol. 1990 Jun;172(6):2911–2919. doi: 10.1128/jb.172.6.2911-2919.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ojanen T., Helander I. M., Haahtela K., Korhonen T. K., Laakso T. Outer Membrane Proteins and Lipopolysaccharides in Pathovars of Xanthomonas campestris. Appl Environ Microbiol. 1993 Dec;59(12):4143–4151. doi: 10.1128/aem.59.12.4143-4151.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Prentki P., Krisch H. M. In vitro insertional mutagenesis with a selectable DNA fragment. Gene. 1984 Sep;29(3):303–313. doi: 10.1016/0378-1119(84)90059-3. [DOI] [PubMed] [Google Scholar]
- Pugsley A. P. The complete general secretory pathway in gram-negative bacteria. Microbiol Rev. 1993 Mar;57(1):50–108. doi: 10.1128/mr.57.1.50-108.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rhen M., Väisänen-Rhen V., Saraste M., Korhonen T. K. Organization of genes expressing the blood-group-M-specific hemagglutinin of Escherichia coli: identification and nucleotide sequence of the M-agglutinin subunit gene. Gene. 1986;49(3):351–360. doi: 10.1016/0378-1119(86)90371-9. [DOI] [PubMed] [Google Scholar]
- Romantschuk M., Bamford D. H. The causal agent of halo blight in bean, Pseudomonas syringae pv. phaseolicola, attaches to stomata via its pili. Microb Pathog. 1986 Apr;1(2):139–148. doi: 10.1016/0882-4010(86)90016-1. [DOI] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schulte R., Bonas U. Expression of the Xanthomonas campestris pv. vesicatoria hrp gene cluster, which determines pathogenicity and hypersensitivity on pepper and tomato, is plant inducible. J Bacteriol. 1992 Feb;174(3):815–823. doi: 10.1128/jb.174.3.815-823.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sohel I., Puente J. L., Murray W. J., Vuopio-Varkila J., Schoolnik G. K. Cloning and characterization of the bundle-forming pilin gene of enteropathogenic Escherichia coli and its distribution in Salmonella serotypes. Mol Microbiol. 1993 Feb;7(4):563–575. doi: 10.1111/j.1365-2958.1993.tb01147.x. [DOI] [PubMed] [Google Scholar]
- Staskawicz B., Dahlbeck D., Keen N., Napoli C. Molecular characterization of cloned avirulence genes from race 0 and race 1 of Pseudomonas syringae pv. glycinea. J Bacteriol. 1987 Dec;169(12):5789–5794. doi: 10.1128/jb.169.12.5789-5794.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Strom M. S., Lory S. Structure-function and biogenesis of the type IV pili. Annu Rev Microbiol. 1993;47:565–596. doi: 10.1146/annurev.mi.47.100193.003025. [DOI] [PubMed] [Google Scholar]
- Virkola R., Parkkinen J., Hacker J., Korhonen T. K. Sialyloligosaccharide chains of laminin as an extracellular matrix target for S fimbriae of Escherichia coli. Infect Immun. 1993 Oct;61(10):4480–4484. doi: 10.1128/iai.61.10.4480-4484.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wilson M., Lindow S. E. Effect of phenotypic plasticity on epiphytic survival and colonization by Pseudomonas syringae. Appl Environ Microbiol. 1993 Feb;59(2):410–416. doi: 10.1128/aem.59.2.410-416.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wilson M., Lindow S. E. Inoculum Density-Dependent Mortality and Colonization of the Phyllosphere by Pseudomonas syringae. Appl Environ Microbiol. 1994 Jul;60(7):2232–2237. doi: 10.1128/aem.60.7.2232-2237.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wu S. S., Kaiser D. Genetic and functional evidence that Type IV pili are required for social gliding motility in Myxococcus xanthus. Mol Microbiol. 1995 Nov;18(3):547–558. doi: 10.1111/j.1365-2958.1995.mmi_18030547.x. [DOI] [PubMed] [Google Scholar]
- de Groot A., Heijnen I., de Cock H., Filloux A., Tommassen J. Characterization of type IV pilus genes in plant growth-promoting Pseudomonas putida WCS358. J Bacteriol. 1994 Feb;176(3):642–650. doi: 10.1128/jb.176.3.642-650.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- van Doorn J., Boonekamp P. M., Oudega B. Partial characterization of fimbriae of Xanthomonas campestris pv. hyacinthi. Mol Plant Microbe Interact. 1994 May-Jun;7(3):334–344. doi: 10.1094/mpmi-7-0334. [DOI] [PubMed] [Google Scholar]