Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1984 Oct;46(1):251–259. doi: 10.1128/iai.46.1.251-259.1984

Cloning and expression of an afimbrial adhesin (AFA-I) responsible for P blood group-independent, mannose-resistant hemagglutination from a pyelonephritic Escherichia coli strain.

A F Labigne-Roussel, D Lark, G Schoolnik, S Falkow
PMCID: PMC261465  PMID: 6148308

Abstract

The uropathogenic Escherichia coli KS52 strain expresses a mannose-resistant hemagglutinin involving an erythrocyte recognition site distinct from the alpha-digalactoside glycosphingolipid receptor identified for the uropathogenic E. coli strains specifying a P adhesin. The KS52 strain showed three major properties. (i) It agglutinated human erythrocytes of all tested blood groups. (ii) Hemagglutinin activity was found both in the supernatant fluid L-broth cultures and in cells grown on L-agar plates. (iii) No fimbriae in organisms grown on L-agar plates were detected by electron microscopy. Whole-cell DNA from the KS52 strain was size fractionated and cloned into the pHC79 cosmid vector. Three recombinant cosmids expressing a mannose-resistant hemagglutination (MRHA) phenotype were characterized and used to subclone the smallest DNA fragment able to confer the same MRHA properties as the parent strain. A 6.7-kilobase chromosomal DNA fragment cloned in pBR322 (pIL14) was shown to be necessary for host-cell MRHA expression and uroepithelial cell adherence. The insert encoded the production of a 16,000-dalton hemagglutinin. This polypeptide could be detected in culture supernatant fluids, in E. coli minicells harboring the pIL14 plasmid, and, by immunoblotting, in the KS52 strain and E. coli whole cells harboring the pIL14 plasmid. No homology was detected by Southern hybridization between the cloned insert and the DNA of the operon responsible for MRHA in the P-specifying, fimbriate strains (pap operon).

Full text

PDF
253

Images in this article

253Image
on p.253
255Image
on p.255
256Image
on p.256
257Image
on p.257

Selected References

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

  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. Cohen M. L., Falkow S. Protein antigens from Staphylococcus aureus strains associated with toxic-shock syndrome. Science. 1981 Feb 20;211(4484):842–844. doi: 10.1126/science.7466361. [DOI] [PubMed] [Google Scholar]
  4. Collins J. Escherichia coli plasmids packageable in vitro in lambda bacteriophage particles. Methods Enzymol. 1979;68:309–326. doi: 10.1016/0076-6879(79)68022-9. [DOI] [PubMed] [Google Scholar]
  5. Davies D. L., Falkiner F. R., Hardy K. G. Colicin V production by clinical isolates of Escherichia coli. Infect Immun. 1981 Feb;31(2):574–579. doi: 10.1128/iai.31.2.574-579.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  7. Duguid J. P., Clegg S., Wilson M. I. The fimbrial and non-fimbrial haemagglutinins of Escherichia coli. J Med Microbiol. 1979 May;12(2):213–227. doi: 10.1099/00222615-12-2-213. [DOI] [PubMed] [Google Scholar]
  8. Edén C. S., Hanson L. A., Jodal U., Lindberg U., Akerlund A. S. Variable adherence to normal human urinary-tract epithelial cells of Escherichia coli strains associated with various forms of urinary-tract infection. Lancet. 1976 Sep 4;1(7984):490–492. [PubMed] [Google Scholar]
  9. Evans D. J., Jr, Evans D. G., Höhne C., Noble M. A., Haldane E. V., Lior H., Young L. S. Hemolysin and K antigens in relation to serotype and hemagglutination type of Escherichia coli isolated from extraintestinal infections. J Clin Microbiol. 1981 Jan;13(1):171–178. doi: 10.1128/jcm.13.1.171-178.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gaastra W., de Graaf F. K. Host-specific fimbrial adhesins of noninvasive enterotoxigenic Escherichia coli strains. Microbiol Rev. 1982 Jun;46(2):129–161. doi: 10.1128/mr.46.2.129-161.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. Hughes C., Hacker J., Roberts A., Goebel W. Hemolysin production as a virulence marker in symptomatic and asymptomatic urinary tract infections caused by Escherichia coli. Infect Immun. 1983 Feb;39(2):546–551. doi: 10.1128/iai.39.2.546-551.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. Ip S. M., Crichton P. B., Old D. C., Duguid J. P. Mannose-resistant and eluting haemagglutinins and fimbriae in Escherichia coli. J Med Microbiol. 1981 May;14(2):223–226. doi: 10.1099/00222615-14-2-223. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. Low D., David V., Lark D., Schoolnik G., Falkow S. Gene clusters governing the production of hemolysin and mannose-resistant hemagglutination are closely linked in Escherichia coli serotype O4 and O6 isolates from urinary tract infections. Infect Immun. 1984 Jan;43(1):353–358. doi: 10.1128/iai.43.1.353-358.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Minshew B. H., Jorgensen J., Counts G. W., Falkow S. Association of hemolysin production, hemagglutination of human erythrocytes, and virulence for chicken embryos of extraintestinal Escherichia coli isolates. Infect Immun. 1978 Apr;20(1):50–54. doi: 10.1128/iai.20.1.50-54.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. 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]
  24. Parkkinen J., Finne J., Achtman M., Väisänen V., Korhonen T. K. Escherichia coli strains binding neuraminyl alpha 2-3 galactosides. Biochem Biophys Res Commun. 1983 Mar 16;111(2):456–461. doi: 10.1016/0006-291x(83)90328-5. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Portnoy D. A., Wolf-Watz H., Bolin I., Beeder A. B., Falkow S. Characterization of common virulence plasmids in Yersinia species and their role in the expression of outer membrane proteins. Infect Immun. 1984 Jan;43(1):108–114. doi: 10.1128/iai.43.1.108-114.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sheladia V. L., Chambers J. P., Guevara J., Jr, Evans D. J. Isolation, purification, and partial characterization of type V-A hemagglutinin from Escherichia coli GV-12, O1:H-. J Bacteriol. 1982 Nov;152(2):757–761. doi: 10.1128/jb.152.2.757-761.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  29. Swaney L. M., Liu Y. P., Ippen-Ihler K., Brinton C. C., Jr Genetic complementation analysis of Escherichia coli type 1 somatic pilus mutants. J Bacteriol. 1977 Apr;130(1):506–511. doi: 10.1128/jb.130.1.506-511.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Väisänen V., Elo J., Tallgren L. G., Siitonen A., Mäkelä P. H., Svanborg-Edén C., Källenius G., Svenson S. B., Hultberg H., Korhonen T. Mannose-resistant haemagglutination and P antigen recognition are characteristic of Escherichia coli causing primary pyelonephritis. Lancet. 1981 Dec 19;2(8260-61):1366–1369. doi: 10.1016/s0140-6736(81)92796-3. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Welch R. A., Dellinger E. P., Minshew B., Falkow S. Haemolysin contributes to virulence of extra-intestinal E. coli infections. Nature. 1981 Dec 17;294(5842):665–667. doi: 10.1038/294665a0. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES