Molecular and General Genetics MGG

, Volume 199, Issue 3, pp 410–414 | Cite as

The fim genes responsible for synthesis of type 1 fimbriae in Escherichia coli, cloning and genetic organization

  • Per Klemm
  • Birthe Jul Jørgensen
  • Irma van Die
  • Han de Ree
  • Hans Bergmans
Article

Summary

The genes responsible for the expression of type 1 fimbriae, produced by the majority of E. coli strains, have been cloned from an E. coli K12 strain. The “passenger” DNA from an initial cosmid clone was reduced in size and subcloned in pACYC184 and pBR322 vectors. A DNA fragment of around 8 kbp was found to be required for the biosynthesis of type 1 fimbriae. This was further studied by transposon-mediated insertional inactivation and by BAL31-mediated deletions. Four genes, designated fimA, B, C, and D were found to be involved in the synthesis of the fimbriae. They encoded proteins that in their processed form appeared with apparent molecular weights of 16.5 kd, 23 kd, 26 kd, and 89 kd, the 16.6 kd polypeptide being the fimbrial subunit. The order to the genes was found to be: fimB, fimA, fimC, and fimD, organized in three transcriptional units.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Adler HI, Fisher WD, Cohon H, Hardigree AA (1967). Miniature Escherichia coli cells deficient in DNA. Proc Natl Acad Sci USA 57:321–326Google Scholar
  2. Andreoli PM, Overbeeke N, Veltkamp E, van Embden JDA, Nijkamp HJJ (1978) Genetic map of the bacteriocinogenic plasmid C10 DF13 derived by insertion of the transposon Tn901. Mol Gen Genet 160:1–11Google Scholar
  3. Bolivar F, Rodriguez RL, Greene PJ, Betlak MC, Heyneker HL, Boyer HW (1977) Construction and characterization of new cloning vehicles II. A multipurpose cloning system. Gene 2:95–113Google Scholar
  4. Boyer HW, Roulland-Dussoix D (1976) A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol 41:459–472Google Scholar
  5. Brinton CC Jr, Gemski P, Falkow S, Baron LS (1961) Location of the piliation factor on the chromosome of Escherichia coli. Biochem Biophys Res Commun 5:293–298Google Scholar
  6. Chang ACY, Cohen SN (1978) Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol 134:1141–1156Google Scholar
  7. Davis RW, Botstein D and Roth (1980) Advanced bacterial genetics. A manual for genetic engineering. Cold Spring Harbor Laboratory, New YorkGoogle Scholar
  8. Die I van, Hondel V van den, Hamstra H-J, Hoekstra W, Bergmans H (1983) Studies on the fimbriae of an Escherichia coli 06:K2:H1:F7 strain: molecular cloning of a DNA fragment encoding a fimbrial antigen responsible for mannose-resistant haemagglutination of human erythrocytes. FEMS Microbiol Lett 19:77–82Google Scholar
  9. Die I van, Megen I van, Hoekstra W, Bergmans H (1984) Molecular organization of the genes involved in the production of a uropathogenic Escherichia coli 06:K2:H1:F7 strain. Mol Gen Genet 194:528–533Google Scholar
  10. Duguid JP, Smith IW, Dempster G, Edmunds PN (1955) Nonflagellar filamentous appendages and hemagglutinating activity in Bacterium coli. J Pathol Bacteriol 70:335–348Google Scholar
  11. Duguid JP, Clegg S, Wilson M (1979) The fimbrial and nonfimbrial hemagglutinins of Escherichia coli. J Med Microbiol 12:213–227Google Scholar
  12. Eisenstein BI (1981) Phase variation of type I fimbriae in Escherichia coli is under transcriptional control. Science 214:337–339Google Scholar
  13. Eshdat Y, Silverblatt FJ, Sharon N (1981) Dissociation and reassembly of Escherichia coli type 1 pili. J Bacteriol 148:308–314Google Scholar
  14. Freitag CS, Eisenstein BI (1983) Genetic mapping and transcriptional orientation of the fimD gene. J Bacteriol 156:1052–1058Google Scholar
  15. Graaf FK de, Krenn BE, Klaasen P (1984) Organization and expression of genes involved in the biosynthesis of K99 fimbriae. Infect Immun 43:508–514Google Scholar
  16. Hagberg L, Jodal U, Korhonen TK, Lindin-Johnson G, Linberg U, Svanborg-Eden C (1981) Adhesion, haemagglutination and virulence of Escherichia coli causing urinary tract infection. Infect Immun 31:564–570Google Scholar
  17. Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:193–197Google Scholar
  18. Iyer R, Darby V, Holland IB (1976) Changes in membrane proteins of Escherichia coli K12 mediated by bacteriophage Ike-specified plasmids. Biochem Biophys Acta 453:311–318Google Scholar
  19. Klemm P (1984) The fimA gene encoding the type 1 fimbrial subunit of Escherichia coli: nucleotide sequence and primary structure of the protein. Eur J Biochem 143:395–399Google Scholar
  20. Klemm P, Ørskov I, Ørskov F (1982) F7 and type 1-like fimbriae from three Escherichia coli strains isolated from urinary tract infection-protein chemical and immunological aspects. Infect Immun 36:462–468Google Scholar
  21. Meyenburg K von, Hansen FG, Nielsen LD, Jørgensen P (1977) Origin of replication, oriC, of the Escherichia coli chromosome: Mapping of genes relative to R. EcoRI cleavage sites in the oriC region. Mol Gen Genet 158:101–109Google Scholar
  22. Meyenburg K von, Hansen FG, Nielsen LD, Riise E (1978) Origin of replication, oriC, of the Escherichia coli chromosome on specialized transducing phages λasn. Mol Gen Genet 160:287–295Google Scholar
  23. Mooi FR, Harms N, Bakker D, Graaf FK de (1981) Organization and expression of genes involved in the production of the K88ab antigen. Infect Immun 32:1155–1163Google Scholar
  24. Mooi FR, Wouters C, Wijfjes A, de Graaf FK (1982) Construction and characterization of mutants impaired in the biosynthesis of the K88 antigen. J Bacteriol 150:512–521Google Scholar
  25. Normark S, Lark D, Hull R, Norgren M, Båga M, O'Hanley P, Schoolnik G, Falkow S (1983) Genetics of digalactoside-binding adhesin from a uropathogenic Escherichia coli strain. Infect Immun 41:942–949Google Scholar
  26. Old DC (1972) Inhibition of interaction between fimbrial haemagglutinins and erythrocytes by D-mannose and other carbohydrates. J Gen Microbiol 71:149–157Google Scholar
  27. Orndorff PE, Falkow S (1984) Organization and expression of genes responcible for type 1 pilliation in Escherichia coli. J Bacteriol 159:736–744Google Scholar
  28. Ørskov I, Ørskov F, Birch-Andersen A (1980) Comparison of Escherichia coli fimbrial antigen F7 with type 1 fimbriae. Infect Immun 27:657–666Google Scholar
  29. Ørskov I, Ørskov F, Birch-Andersen A, Klemm P, Svanborg-Eden C (1982) Protein attachment factors: Fimbriae in adhering Escherichia coli strains, Seminars in infectious disease, vol IV, bacterial vaccines. Thieme-Stratton Inc., New York, pp 97–103Google Scholar
  30. Swaney LM, Liu Y-P, To CM, To CC, Ippen-Ihler K, Brinton Jr CC (1977a) Isolation and characterization of Escherichia coli phase variants and mutants deficient in type 1 pilus production. J Bacteriol 130:495–505Google Scholar
  31. Swaney LM, Liu Y-P, Ippen-Ihler K, Brinton Jr CC (1977b) Genetic complementation analysis of Escherichia coli type 1 somatic pilus mutants. J Bacteriol 130:506–511Google Scholar
  32. Timmis K, Cabello F, Cohen SN (1978) Cloning and characterization of EcoRI and Hind III restriction endonuclease-generated fragments of antibiotic resistance plasmids. Mol Gen Genet 162:121–137Google Scholar

Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • Per Klemm
    • 1
  • Birthe Jul Jørgensen
    • 1
  • Irma van Die
    • 2
  • Han de Ree
    • 2
  • Hans Bergmans
    • 2
  1. 1.Department of MicrobiologyThe Technical University of DenmarkLyngbyDenmark
  2. 2.Department of Molecular Cell BiologyState University of UtrechtUtrechtThe Netherlands

Personalised recommendations