Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1997 Jun;179(11):3594–3603. doi: 10.1128/jb.179.11.3594-3603.1997

The plasmid R64 thin pilus identified as a type IV pilus.

S R Kim 1, T Komano 1
PMCID: PMC179153  PMID: 9171405

Abstract

The entire nucleotide sequence of the pil region of the IncI1 plasmid R64 was determined. Analysis of the sequence indicated that 14 genes, designated pilI through pilV, are involved in the formation of the R64 thin pilus. Protein products of eight pil genes were identified by the maxicell procedure. The pilN product was shown to be a lipoprotein by an experiment using globomycin. A computer search revealed that several R64 pil genes have amino acid sequence homology with proteins involved in type IV pilus biogenesis, protein secretion, and transformation competence. The pilS and pilV products were suggested to be prepilins for the R64 thin pilus, and the pilU product appears to be a prepilin peptidase. These results suggest that the R64 thin pilus belongs to the type IV family, specifically group IVB, of pili. The requirement of the pilR and pilU genes for R64 liquid mating was demonstrated by constructing their frameshift mutations. Comparison of three type IVB pilus biogenesis systems, the pil system of R64, the toxin-coregulated pilus (tcp) system of Vibrio cholerae, and the bundle-forming pilus (bfp) system of enteropathogenic Escherichia coli, suggests that they have evolved from a common ancestral gene system.

Full Text

The Full Text of this article is available as a PDF (936.7 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allaoui A., Ménard R., Sansonetti P. J., Parsot C. Characterization of the Shigella flexneri ipgD and ipgF genes, which are located in the proximal part of the mxi locus. Infect Immun. 1993 May;61(5):1707–1714. doi: 10.1128/iai.61.5.1707-1714.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bayer M., Eferl R., Zellnig G., Teferle K., Dijkstra A., Koraimann G., Högenauer G. Gene 19 of plasmid R1 is required for both efficient conjugative DNA transfer and bacteriophage R17 infection. J Bacteriol. 1995 Aug;177(15):4279–4288. doi: 10.1128/jb.177.15.4279-4288.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradley D. E. Characteristics and function of thick and thin conjugative pili determined by transfer-derepressed plasmids of incompatibility groups I1, I2, I5, B, K and Z. J Gen Microbiol. 1984 Jun;130(6):1489–1502. doi: 10.1099/00221287-130-6-1489. [DOI] [PubMed] [Google Scholar]
  4. Bradley D. E. Derepressed plasmids of incompatibility group I1 determine two different morphological forms of pilus. Plasmid. 1983 May;9(3):331–334. doi: 10.1016/0147-619x(83)90011-2. [DOI] [PubMed] [Google Scholar]
  5. Coetzee J. N., Bradley D. E., Hedges R. W. Phages I alpha and I2-2: IncI plasmid-dependent bacteriophages. J Gen Microbiol. 1982 Nov;128(11):2797–2804. doi: 10.1099/00221287-128-11-2797. [DOI] [PubMed] [Google Scholar]
  6. Coetzee J. N., Sirgel F. A., Lecatsas G. Properties of a filamentous phage which adsorbs to pili coded by plasmids of the IncI complex. J Gen Microbiol. 1980 Apr;117(2):547–551. doi: 10.1099/00221287-117-2-547. [DOI] [PubMed] [Google Scholar]
  7. DiRita V. J., Parsot C., Jander G., Mekalanos J. J. Regulatory cascade controls virulence in Vibrio cholerae. Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5403–5407. doi: 10.1073/pnas.88.12.5403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dijkstra A. J., Keck W. Peptidoglycan as a barrier to transenvelope transport. J Bacteriol. 1996 Oct;178(19):5555–5562. doi: 10.1128/jb.178.19.5555-5562.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dijkstra B. W., Thunnissen A. M. 'Holy' proteins. II: The soluble lytic transglycosylase. Curr Opin Struct Biol. 1994 Dec;4(6):810–813. doi: 10.1016/0959-440x(94)90261-5. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Girón J. A., Levine M. M., Kaper J. B. Longus: a long pilus ultrastructure produced by human enterotoxigenic Escherichia coli. Mol Microbiol. 1994 Apr;12(1):71–82. doi: 10.1111/j.1365-2958.1994.tb00996.x. [DOI] [PubMed] [Google Scholar]
  12. Hobbs M., Mattick J. S. Common components in the assembly of type 4 fimbriae, DNA transfer systems, filamentous phage and protein-secretion apparatus: a general system for the formation of surface-associated protein complexes. Mol Microbiol. 1993 Oct;10(2):233–243. doi: 10.1111/j.1365-2958.1993.tb01949.x. [DOI] [PubMed] [Google Scholar]
  13. Hultgren S. J., Abraham S., Caparon M., Falk P., St Geme J. W., 3rd, Normark S. Pilus and nonpilus bacterial adhesins: assembly and function in cell recognition. Cell. 1993 Jun 4;73(5):887–901. doi: 10.1016/0092-8674(93)90269-v. [DOI] [PubMed] [Google Scholar]
  14. Inukai M., Takeuchi M., Shimizu K., Arai M. Mechanism of action of globomycin. J Antibiot (Tokyo) 1978 Nov;31(11):1203–1205. doi: 10.7164/antibiotics.31.1203. [DOI] [PubMed] [Google Scholar]
  15. Kim S. R., Funayama N., Komano T. Nucleotide sequence and characterization of the traABCD region of IncI1 plasmid R64. J Bacteriol. 1993 Aug;175(16):5035–5042. doi: 10.1128/jb.175.16.5035-5042.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kim S. R., Komano T. Nucleotide sequence of the R721 shufflon. J Bacteriol. 1992 Nov;174(21):7053–7058. doi: 10.1128/jb.174.21.7053-7058.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Komano T., Funayama N., Kim S. R., Nisioka T. Transfer region of IncI1 plasmid R64 and role of shufflon in R64 transfer. J Bacteriol. 1990 May;172(5):2230–2235. doi: 10.1128/jb.172.5.2230-2235.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Komano T., Kim S. R., Yoshida T. Mating variation by DNA inversions of shufflon in plasmid R64. Adv Biophys. 1995;31:181–193. doi: 10.1016/0065-227x(95)99391-2. [DOI] [PubMed] [Google Scholar]
  19. Komano T., Kim S. R., Yoshida T., Nisioka T. DNA rearrangement of the shufflon determines recipient specificity in liquid mating of IncI1 plasmid R64. J Mol Biol. 1994 Oct 14;243(1):6–9. doi: 10.1006/jmbi.1994.1625. [DOI] [PubMed] [Google Scholar]
  20. Komano T., Kubo A., Nisioka T. Shufflon: multi-inversion of four contiguous DNA segments of plasmid R64 creates seven different open reading frames. Nucleic Acids Res. 1987 Feb 11;15(3):1165–1172. doi: 10.1093/nar/15.3.1165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kubo A., Kusukawa A., Komano T. Nucleotide sequence of the rci gene encoding shufflon-specific DNA recombinase in the IncI1 plasmid R64: homology to the site-specific recombinases of integrase family. Mol Gen Genet. 1988 Jul;213(1):30–35. doi: 10.1007/BF00333394. [DOI] [PubMed] [Google Scholar]
  22. Kuehn M. J., Heuser J., Normark S., Hultgren S. J. P pili in uropathogenic E. coli are composite fibres with distinct fibrillar adhesive tips. Nature. 1992 Mar 19;356(6366):252–255. doi: 10.1038/356252a0. [DOI] [PubMed] [Google Scholar]
  23. Lindberg F., Lund B., Johansson L., Normark S. Localization of the receptor-binding protein adhesin at the tip of the bacterial pilus. Nature. 1987 Jul 2;328(6125):84–87. doi: 10.1038/328084a0. [DOI] [PubMed] [Google Scholar]
  24. Miras I., Hermant D., Arricau N., Popoff M. Y. Nucleotide sequence of iagA and iagB genes involved in invasion of HeLa cells by Salmonella enterica subsp. enterica ser. Typhi. Res Microbiol. 1995 Jan;146(1):17–20. doi: 10.1016/0923-2508(96)80267-1. [DOI] [PubMed] [Google Scholar]
  25. Ogierman M. A., Zabihi S., Mourtzios L., Manning P. A. Genetic organization and sequence of the promoter-distal region of the tcp gene cluster of Vibrio cholerae. Gene. 1993 Apr 15;126(1):51–60. doi: 10.1016/0378-1119(93)90589-u. [DOI] [PubMed] [Google Scholar]
  26. Ottow J. C. Ecology, physiology, and genetics of fimbriae and pili. Annu Rev Microbiol. 1975;29:79–108. doi: 10.1146/annurev.mi.29.100175.000455. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Rudel T., Scheurerpflug I., Meyer T. F. Neisseria PilC protein identified as type-4 pilus tip-located adhesin. Nature. 1995 Jan 26;373(6512):357–359. doi: 10.1038/373357a0. [DOI] [PubMed] [Google Scholar]
  29. Sancar A., Hack A. M., Rupp W. D. Simple method for identification of plasmid-coded proteins. J Bacteriol. 1979 Jan;137(1):692–693. doi: 10.1128/jb.137.1.692-693.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Sohel I., Puente J. L., Ramer S. W., Bieber D., Wu C. Y., Schoolnik G. K. Enteropathogenic Escherichia coli: identification of a gene cluster coding for bundle-forming pilus morphogenesis. J Bacteriol. 1996 May;178(9):2613–2628. doi: 10.1128/jb.178.9.2613-2628.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Stone K. D., Zhang H. Z., Carlson L. K., Donnenberg M. S. A cluster of fourteen genes from enteropathogenic Escherichia coli is sufficient for the biogenesis of a type IV pilus. Mol Microbiol. 1996 Apr;20(2):325–337. doi: 10.1111/j.1365-2958.1996.tb02620.x. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Strom M. S., Nunn D. N., Lory S. A single bifunctional enzyme, PilD, catalyzes cleavage and N-methylation of proteins belonging to the type IV pilin family. Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2404–2408. doi: 10.1073/pnas.90.6.2404. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Thunnissen A. M., Dijkstra A. J., Kalk K. H., Rozeboom H. J., Engel H., Keck W., Dijkstra B. W. Doughnut-shaped structure of a bacterial muramidase revealed by X-ray crystallography. Nature. 1994 Feb 24;367(6465):750–753. doi: 10.1038/367750a0. [DOI] [PubMed] [Google Scholar]
  36. Vieira J., Messing J. Production of single-stranded plasmid DNA. Methods Enzymol. 1987;153:3–11. doi: 10.1016/0076-6879(87)53044-0. [DOI] [PubMed] [Google Scholar]
  37. Walker J. E., Saraste M., Runswick M. J., Gay N. J. Distantly related sequences in the alpha- and beta-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold. EMBO J. 1982;1(8):945–951. doi: 10.1002/j.1460-2075.1982.tb01276.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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]
  39. 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]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES