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. 1985 Jun;162(3):1285–1292. doi: 10.1128/jb.162.3.1285-1292.1985

Genetic organization of the afimbrial adhesin operon and nucleotide sequence from a uropathogenic Escherichia coli gene encoding an afimbrial adhesin.

A Labigne-Roussel, M A Schmidt, W Walz, S Falkow
PMCID: PMC215917  PMID: 2860096

Abstract

The uropathogenic Escherichia coli KS52 strain expresses a mannose-resistant hemagglutinin AFA-I, which recognizes a human erythrocyte site distinct from the alpha-digalactoside glycosphingolipid receptor common to uropathogenic E. coli strains specifying a P adhesin. A 6.7-kilobase chromosomal DNA fragment was cloned from KS52 into pBR322 and was shown to be necessary for host cell mannose-resistant hemagglutination expression and uroepithelial cell adherence (Labigne-Roussel et al., Infect. Immun. 46:251-259, 1984). The genetic organization of the 6.7-kilobase DNA fragment was investigated by generating derivative plasmids, and the polypeptides encoded by those plasmids in isolated minicells were analyzed on polyacrylamide gel. The 6.7-kilobase insert expresses five polypeptides of molecular mass 13,000, 16,000, 18,500, 30,000, and 100,000 daltons encoded, respectively, by the afaA, afaE, afaD, afaB, and afaC genes. The five genes were localized and were shown to belong to the same transcriptional unit. The afaB, afaC, and afaE gene products are required for mannose-resistant hemagglutination expression, whereas mutations in or deletions of either afaA or afaD do not modify host mannose-resistant hemagglutination expression. The afaE gene was identified as the structural gene encoding hemagglutinin. The gene has been sequenced; it encodes a 152-residue protein containing a typical 21-residue procaryote signal sequence and a 131-residue mature polypeptide, the AFA-I adhesin.

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Selected References

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  1. Adler H. I., Fisher W. D., Cohen A., Hardigree A. A. MINIATURE escherichia coli CELLS DEFICIENT IN DNA. Proc Natl Acad Sci U S A. 1967 Feb;57(2):321–326. doi: 10.1073/pnas.57.2.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
  3. Dallas W. S., Falkow S. The molecular nature of heat-labile enterotoxin (LT) of escherichia coli. Nature. 1979 Feb 1;277(5695):406–407. doi: 10.1038/277406a0. [DOI] [PubMed] [Google Scholar]
  4. Dougan G., Saul M., Twigg A., Gill R., Sherratt D. Polypeptides expressed in Escherichia coli K-12 minicells by transposition elements Tn1 and Tn3. J Bacteriol. 1979 Apr;138(1):48–54. doi: 10.1128/jb.138.1.48-54.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gill R. E., Heffron F., Falkow S. Identification of the protein encoded by the transposable element Tn3 which is required for its transposition. Nature. 1979 Dec 20;282(5741):797–801. doi: 10.1038/282797a0. [DOI] [PubMed] [Google Scholar]
  6. Gold L., Pribnow D., Schneider T., Shinedling S., Singer B. S., Stormo G. Translational initiation in prokaryotes. Annu Rev Microbiol. 1981;35:365–403. doi: 10.1146/annurev.mi.35.100181.002053. [DOI] [PubMed] [Google Scholar]
  7. Hagberg L., Jodal U., Korhonen T. K., Lidin-Janson G., Lindberg U., Svanborg Edén C. Adhesion, hemagglutination, and virulence of Escherichia coli causing urinary tract infections. Infect Immun. 1981 Feb;31(2):564–570. doi: 10.1128/iai.31.2.564-570.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hughes C., Phillips R., Roberts A. P. Serum resistance among Escherichia coli strains causing urinary tract infection in relation to O type and the carriage of hemolysin, colicin, and antibiotic resistance determinants. Infect Immun. 1982 Jan;35(1):270–275. doi: 10.1128/iai.35.1.270-275.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hull R. A., Gill R. E., Hsu P., Minshew B. H., Falkow S. Construction and expression of recombinant plasmids encoding type 1 or D-mannose-resistant pili from a urinary tract infection Escherichia coli isolate. Infect Immun. 1981 Sep;33(3):933–938. doi: 10.1128/iai.33.3.933-938.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Inouye M., Halegoua S. Secretion and membrane localization of proteins in Escherichia coli. CRC Crit Rev Biochem. 1980;7(4):339–371. doi: 10.3109/10409238009105465. [DOI] [PubMed] [Google Scholar]
  11. Källenius G., Svenson S. B., Möllby R., Korhonen T., Winberg J., Cedergren B., Helin I., Hultberg H. Carbohydrate receptor structures recognized by uropathogenic E. coli. Scand J Infect Dis Suppl. 1982;33:52–60. [PubMed] [Google Scholar]
  12. Källenius G., Svenson S., Möllby R., Cedergren B., Hultberg H., Winberg J. Structure of carbohydrate part of receptor on human uroepithelial cells for pyelonephritogenic Escherichia coli. Lancet. 1981 Sep 19;2(8247):604–606. doi: 10.1016/s0140-6736(81)92743-4. [DOI] [PubMed] [Google Scholar]
  13. Labigne-Roussel A. F., Lark D., Schoolnik G., Falkow S. Cloning and expression of an afimbrial adhesin (AFA-I) responsible for P blood group-independent, mannose-resistant hemagglutination from a pyelonephritic Escherichia coli strain. Infect Immun. 1984 Oct;46(1):251–259. doi: 10.1128/iai.46.1.251-259.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lindberg F. P., Lund B., Normark S. Genes of pyelonephritogenic E. coli required for digalactoside-specific agglutination of human cells. EMBO J. 1984 May;3(5):1167–1173. doi: 10.1002/j.1460-2075.1984.tb01946.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Messing J., Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. doi: 10.1016/0378-1119(82)90016-6. [DOI] [PubMed] [Google Scholar]
  16. Mooi F. R., van Buuren M., Koopman G., Roosendaal B., de Graaf F. K. K88ab gene of Escherichia coli encodes a fimbria-like protein distinct from the K88ab fimbrial adhesin. J Bacteriol. 1984 Aug;159(2):482–487. doi: 10.1128/jb.159.2.482-487.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Norgren M., Normark S., Lark D., O'Hanley P., Schoolnik G., Falkow S., Svanborg-Edén C., Båga M., Uhlin B. E. Mutations in E coli cistrons affecting adhesion to human cells do not abolish Pap pili fiber formation. EMBO J. 1984 May;3(5):1159–1165. doi: 10.1002/j.1460-2075.1984.tb01945.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Normark S., Lark D., Hull R., Norgren M., Båga M., O'Hanley P., Schoolnik G., Falkow S. Genetics of digalactoside-binding adhesin from a uropathogenic Escherichia coli strain. Infect Immun. 1983 Sep;41(3):942–949. doi: 10.1128/iai.41.3.942-949.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. O'Hanley P., Lark D., Normark S., Falkow S., Schoolnik G. K. Mannose-sensitive and Gal-Gal binding Escherichia coli pili from recombinant strains. Chemical, functional, and serological properties. J Exp Med. 1983 Nov 1;158(5):1713–1719. doi: 10.1084/jem.158.5.1713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Palva E. T., Hirst T. R., Hardy S. J., Holmgren J., Randall L. Synthesis of a precursor to the B subunit of heat-labile enterotoxin in Escherichia coli. J Bacteriol. 1981 Apr;146(1):325–330. doi: 10.1128/jb.146.1.325-330.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Portnoy D. A., Moseley S. L., Falkow S. Characterization of plasmids and plasmid-associated determinants of Yersinia enterocolitica pathogenesis. Infect Immun. 1981 Feb;31(2):775–782. doi: 10.1128/iai.31.2.775-782.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sanger F., Coulson A. R., Barrell B. G., Smith A. J., Roe B. A. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol. 1980 Oct 25;143(2):161–178. doi: 10.1016/0022-2836(80)90196-5. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Stormo G. D., Schneider T. D., Gold L. M. Characterization of translational initiation sites in E. coli. Nucleic Acids Res. 1982 May 11;10(9):2971–2996. doi: 10.1093/nar/10.9.2971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Väisänen V., Korhonen T. K., Jokinen M., Gahmberg C. G., Ehnholm C. Blood group M specific haemagglutinin in pyelonephritogenic Escherichia coli. Lancet. 1982 May 22;1(8282):1192–1192. doi: 10.1016/s0140-6736(82)92264-4. [DOI] [PubMed] [Google Scholar]
  26. 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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