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. 1995 Feb;63(2):724–728. doi: 10.1128/iai.63.2.724-728.1995

Sequencing of the gene encoding the major pilin of pilus colonization factor antigen III (CFA/III) of human enterotoxigenic Escherichia coli and evidence that CFA/III is related to type IV pili.

T Taniguchi 1, Y Fujino 1, K Yamamoto 1, T Miwatani 1, T Honda 1
PMCID: PMC173060  PMID: 7822050

Abstract

The plasmid-encoded structural gene cofA necessary for the production of the major pilin subunit of pilus colonization factor antigen III (CFA/III) of human enterotoxigenic Escherichia coli was identified, and the nucleotide sequence of the gene was determined. cofA consists of 714 nucleotides encoding a 238-amino-acid protein (molecular weight of 25,309). CofA seems to be a precursor of CFA/III pilin, because the first 23 residues of the N-terminal amino acid sequence of the purified CFA/III pili coincided with the deduced amino acid sequence for residues 32 to 54 of CofA. Western blot (immunoblot) analysis of CofA also indicated its processing to form mature pilin in the presence of the downstream region of cofA. These results suggest that the major pilin of CFA/III pili is produced as a precursor form which is posttranslationally modified to the mature pilin and forms morphological pili after cleavage of the Gly-30-Met-31 junction, probably by a protease encoded by an as-yet-unknown gene located downstream of cofA. Interestingly, the N-terminal 30-amino-acid sequence of mature CFA/III shows the highest identity (76.7%) to TcpA pilin of Vibrio cholerae, which is a type IV class B pilin.

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Selected References

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  1. Bakker D., Vader C. E., Roosendaal B., Mooi F. R., Oudega B., de Graaf F. K. Structure and function of periplasmic chaperone-like proteins involved in the biosynthesis of K88 and K99 fimbriae in enterotoxigenic Escherichia coli. Mol Microbiol. 1991 Apr;5(4):875–886. doi: 10.1111/j.1365-2958.1991.tb00761.x. [DOI] [PubMed] [Google Scholar]
  2. Boylan M., Smyth C. J., Scott J. R. Nucleotide sequence of the gene encoding the major subunit of CS3 fimbriae of enterotoxigenic Escherichia coli. Infect Immun. 1988 Dec;56(12):3297–3300. doi: 10.1128/iai.56.12.3297-3300.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clark C. A., Heuzenroeder M. W., Manning P. A. Colonization factor antigen CFA/IV (PCF8775) of human enterotoxigenic Escherichia coli: nucleotide sequence of the CS5 determinant. Infect Immun. 1992 Mar;60(3):1254–1257. doi: 10.1128/iai.60.3.1254-1257.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cravioto A., Scotland S. M., Rowe B. Hemagglutination activity and colonization factor antigens I and II in enterotoxigenic and non-enterotoxigenic strains of Escherichia coli isolated from humans. Infect Immun. 1982 Apr;36(1):189–197. doi: 10.1128/iai.36.1.189-197.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Elleman T. C., Hoyne P. A., McKern N. M., Stewart D. J. Nucleotide sequence of the gene encoding the two-subunit pilin of Bacteroides nodosus 265. J Bacteriol. 1986 Jul;167(1):243–250. doi: 10.1128/jb.167.1.243-250.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Evans D. G., Evans D. J., Jr, Tjoa W. Hemagglutination of human group A erythrocytes by enterotoxigenic Escherichia coli isolated from adults with diarrhea: correlation with colonization factor. Infect Immun. 1977 Nov;18(2):330–337. doi: 10.1128/iai.18.2.330-337.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Evans D. G., Silver R. P., Evans D. J., Jr, Chase D. G., Gorbach S. L. Plasmid-controlled colonization factor associated with virulence in Esherichia coli enterotoxigenic for humans. Infect Immun. 1975 Sep;12(3):656–667. doi: 10.1128/iai.12.3.656-667.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Faast R., Ogierman M. A., Stroeher U. H., Manning P. A. Nucleotide sequence of the structural gene, tcpA, for a major pilin subunit of Vibrio cholerae. Gene. 1989 Dec 21;85(1):227–231. doi: 10.1016/0378-1119(89)90486-1. [DOI] [PubMed] [Google Scholar]
  11. Fulks K. A., Marrs C. F., Stevens S. P., Green M. R. Sequence analysis of the inversion region containing the pilin genes of Moraxella bovis. J Bacteriol. 1990 Jan;172(1):310–316. doi: 10.1128/jb.172.1.310-316.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Guth B. E., Aguiar E. G., Griffin P. M., Ramos S. R., Gomes T. A. Prevalence of colonization factor antigens (CFAs) and adherence to HeLa cells in enterotoxigenic Escherichia coli isolated from feces of children in São Paulo. Microbiol Immunol. 1994;38(9):695–701. doi: 10.1111/j.1348-0421.1994.tb01844.x. [DOI] [PubMed] [Google Scholar]
  14. Hamers A. M., Pel H. J., Willshaw G. A., Kusters J. G., van der Zeijst B. A., Gaastra W. The nucleotide sequence of the first two genes of the CFA/I fimbrial operon of human enterotoxigenic Escherichia coli. Microb Pathog. 1989 Apr;6(4):297–309. doi: 10.1016/0882-4010(89)90103-4. [DOI] [PubMed] [Google Scholar]
  15. Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
  16. Heuzenroeder M. W., Neal B. L., Thomas C. J., Halter R., Manning P. A. Characterization and molecular cloning of the PCF8775 CS5 antigen from an enterotoxigenic Escherichia coli 0115:H40 isolated in Central Australia. Mol Microbiol. 1989 Mar;3(3):303–310. doi: 10.1111/j.1365-2958.1989.tb00175.x. [DOI] [PubMed] [Google Scholar]
  17. Hobbs M., Dalrymple B. P., Cox P. T., Livingstone S. P., Delaney S. F., Mattick J. S. Organization of the fimbrial gene region of Bacteroides nodosus: class I and class II strains. Mol Microbiol. 1991 Mar;5(3):543–560. doi: 10.1111/j.1365-2958.1991.tb00726.x. [DOI] [PubMed] [Google Scholar]
  18. Honda T., Arita M., Miwatani T. Characterization of new hydrophobic pili of human enterotoxigenic Escherichia coli: a possible new colonization factor. Infect Immun. 1984 Mar;43(3):959–965. doi: 10.1128/iai.43.3.959-965.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Honda T., Wetprasit N., Arita M., Miwatani T. Production and characterization of monoclonal antibodies to a pilus colonization factor (colonization factor antigen III) of human enterotoxigenic Escherichia coli. Infect Immun. 1989 Nov;57(11):3452–3457. doi: 10.1128/iai.57.11.3452-3457.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Karjalainen T. K., Evans D. G., So M., Lee C. H. Molecular cloning and nucleotide sequence of the colonization factor antigen I gene of Escherichia coli. Infect Immun. 1989 Apr;57(4):1126–1130. doi: 10.1128/iai.57.4.1126-1130.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Levine M. M., Ristaino P., Marley G., Smyth C., Knutton S., Boedeker E., Black R., Young C., Clements M. L., Cheney C. Coli surface antigens 1 and 3 of colonization factor antigen II-positive enterotoxigenic Escherichia coli: morphology, purification, and immune responses in humans. Infect Immun. 1984 May;44(2):409–420. doi: 10.1128/iai.44.2.409-420.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. McConnell M. M., Thomas L. V., Willshaw G. A., Smith H. R., Rowe B. Genetic control and properties of coli surface antigens of colonization factor antigen IV (PCF8775) of enterotoxigenic Escherichia coli. Infect Immun. 1988 Aug;56(8):1974–1980. doi: 10.1128/iai.56.8.1974-1980.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Meyer T. F., Billyard E., Haas R., Storzbach S., So M. Pilus genes of Neisseria gonorrheae: chromosomal organization and DNA sequence. Proc Natl Acad Sci U S A. 1984 Oct;81(19):6110–6114. doi: 10.1073/pnas.81.19.6110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mullany P., Field A. M., McConnell M. M., Scotland S. M., Smith H. R., Rowe B. Expression of plasmids coding for colonization factor antigen II (CFA/II) and enterotoxin production in Escherichia coli. J Gen Microbiol. 1983 Dec;129(12):3591–3601. doi: 10.1099/00221287-129-12-3591. [DOI] [PubMed] [Google Scholar]
  26. Pasloske B. L., Finlay B. B., Paranchych W. Cloning and sequencing of the Pseudomonas aeruginosa PAK pilin gene. FEBS Lett. 1985 Apr 22;183(2):408–412. doi: 10.1016/0014-5793(85)80821-8. [DOI] [PubMed] [Google Scholar]
  27. Pedersen P. A., Andersen L. N. Deletion and duplication of specific sequences in the K88ab fimbrial subunit protein from porcine enterotoxigenic Escherichia coli. Mol Gen Genet. 1991 Oct;229(2):285–291. doi: 10.1007/BF00272168. [DOI] [PubMed] [Google Scholar]
  28. Pedersen P. A. Structure-function analysis of the K88ab fimbrial subunit protein from porcine enterotoxigenic Escherichia coli. Mol Microbiol. 1991 May;5(5):1073–1080. doi: 10.1111/j.1365-2958.1991.tb01879.x. [DOI] [PubMed] [Google Scholar]
  29. Perez-Casal J., Swartley J. S., Scott J. R. Gene encoding the major subunit of CS1 pili of human enterotoxigenic Escherichia coli. Infect Immun. 1990 Nov;58(11):3594–3600. doi: 10.1128/iai.58.11.3594-3600.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Shaw C. E., Taylor R. K. Vibrio cholerae O395 tcpA pilin gene sequence and comparison of predicted protein structural features to those of type 4 pilins. Infect Immun. 1990 Sep;58(9):3042–3049. doi: 10.1128/iai.58.9.3042-3049.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Shinagawa H., Taniguchi T., Yamaguchi O., Yamamoto K., Honda T. Cloning of the genes that control formation of the fimbrial colonization factor antigen III (CFA/III) from an enterotoxigenic Escherichia coli. Microbiol Immunol. 1993;37(9):689–694. doi: 10.1111/j.1348-0421.1993.tb01693.x. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. Smyth C. J. Two mannose-resistant haemagglutinins on enterotoxigenic Escherichia coli of serotype O6:K15:H16 or H-isolated from travellers' and infantile diarrhoea. J Gen Microbiol. 1982 Sep;128(9):2081–2096. doi: 10.1099/00221287-128-9-2081. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. Taniguchi T., Arita M., Sato M., Yamamoto K., Miwatani T., Honda T. Evidence that the N-terminal amino acid sequence of pilus colonization factor antigen III produced by human enterotoxigenic Escherichia coli is similar to that of TcpA pilin of Vibrio cholerae. J Infect Dis. 1994 Oct;170(4):1049–1050. doi: 10.1093/infdis/170.4.1049. [DOI] [PubMed] [Google Scholar]
  38. Thomas L. V., McConnell M. M., Rowe B., Field A. M. The possession of three novel coli surface antigens by enterotoxigenic Escherichia coli strains positive for the putative colonization factor PCF8775. J Gen Microbiol. 1985 Sep;131(9):2319–2326. doi: 10.1099/00221287-131-9-2319. [DOI] [PubMed] [Google Scholar]
  39. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Yamaguchi K., Masamune Y. Autogenous regulation of synthesis of the replication protein in plasmid pSC101. Mol Gen Genet. 1985;200(3):362–367. doi: 10.1007/BF00425718. [DOI] [PubMed] [Google Scholar]
  41. 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]
  42. de Graaf F. K., Krenn B. E., Klaasen P. Organization and expression of genes involved in the biosynthesis of K99 fimbriae. Infect Immun. 1984 Feb;43(2):508–514. doi: 10.1128/iai.43.2.508-514.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]

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