Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1997 Oct;65(10):4135–4145. doi: 10.1128/iai.65.10.4135-4145.1997

Aggregative adherence fimbria II, a second fimbrial antigen mediating aggregative adherence in enteroaggregative Escherichia coli.

J R Czeczulin 1, S Balepur 1, S Hicks 1, A Phillips 1, R Hall 1, M H Kothary 1, F Navarro-Garcia 1, J P Nataro 1
PMCID: PMC175595  PMID: 9317019

Abstract

Enteroaggregative Escherichia coli (EAEC) has been implicated as an agent of pediatric diarrhea in the developing world. We have shown previously that EAEC adheres to HEp-2 cells by virtue of a plasmid-encoded fimbrial adhesin designated aggregative adherence fimbria I (AAF/I), the genes for which have been cloned and sequenced. However, not all EAEC strains express AAF/I. Using TnphoA mutagenesis, we have characterized a novel fimbria (designated AAF/II) which mediates HEp-2 adherence of the human-pathogenic strain 042. AAF/II is 5 nm in diameter and does not bind AAF/I antiserum, as determined by immunogold transmission electron microscopy. TnphoA identified a gene (designated aafA) which bears significant homology to aggA, the fimbrial subunit of AAF/I (25% identity and 47% similarity at the amino acid level). When hyperexpressed and purified by polyhistidine tagging, the AafA protein assembled into 5-nm-diameter filaments which bound anti-AAF/II antiserum. The cloned aafA gene complemented a mutation in the aggA gene to confer fimbrial expression from the AAF/I gene cluster, manifesting phenotypes characteristic of AAF/II but not AAF/I. The aafA mutant did not adhere to human intestinal tissue in culture, suggesting a role for AAF/II in intestinal colonization. By using DNA probes for AAF/I and AAF/II derived from fimbrial biosynthesis genes, we show that AAF/I and AAF/II are each found in only a minority of EAEC strains, suggesting that still more EAEC adhesins exist. Our data suggest that AAF adhesins represent a new family of fimbrial adhesins which mediate aggregative adherence in EAEC.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

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

  1. Baldwin T. J., Knutton S., Sellers L., Hernandez H. A., Aitken A., Williams P. H. Enteroaggregative Escherichia coli strains secrete a heat-labile toxin antigenically related to E. coli hemolysin. Infect Immun. 1992 May;60(5):2092–2095. doi: 10.1128/iai.60.5.2092-2095.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baudry B., Savarino S. J., Vial P., Kaper J. B., Levine M. M. A sensitive and specific DNA probe to identify enteroaggregative Escherichia coli, a recently discovered diarrheal pathogen. J Infect Dis. 1990 Jun;161(6):1249–1251. doi: 10.1093/infdis/161.6.1249. [DOI] [PubMed] [Google Scholar]
  3. Bhan M. K., Khoshoo V., Sommerfelt H., Raj P., Sazawal S., Srivastava R. Enteroaggregative Escherichia coli and Salmonella associated with nondysenteric persistent diarrhea. Pediatr Infect Dis J. 1989 Aug;8(8):499–502. doi: 10.1097/00006454-198908000-00005. [DOI] [PubMed] [Google Scholar]
  4. Bhan M. K., Raj P., Levine M. M., Kaper J. B., Bhandari N., Srivastava R., Kumar R., Sazawal S. Enteroaggregative Escherichia coli associated with persistent diarrhea in a cohort of rural children in India. J Infect Dis. 1989 Jun;159(6):1061–1064. doi: 10.1093/infdis/159.6.1061. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Cravioto A., Tello A., Navarro A., Ruiz J., Villafán H., Uribe F., Eslava C. Association of Escherichia coli HEp-2 adherence patterns with type and duration of diarrhoea. Lancet. 1991 Feb 2;337(8736):262–264. doi: 10.1016/0140-6736(91)90868-p. [DOI] [PubMed] [Google Scholar]
  7. Gicquelais K. G., Baldini M. M., Martinez J., Maggi L., Martin W. C., Prado V., Kaper J. B., Levine M. M. Practical and economical method for using biotinylated DNA probes with bacterial colony blots to identify diarrhea-causing Escherichia coli. J Clin Microbiol. 1990 Nov;28(11):2485–2490. doi: 10.1128/jcm.28.11.2485-2490.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  9. Hicks S., Candy D. C., Phillips A. D. Adhesion of enteroaggregative Escherichia coli to pediatric intestinal mucosa in vitro. Infect Immun. 1996 Nov;64(11):4751–4760. doi: 10.1128/iai.64.11.4751-4760.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Izard J. W., Kendall D. A. Signal peptides: exquisitely designed transport promoters. Mol Microbiol. 1994 Sep;13(5):765–773. doi: 10.1111/j.1365-2958.1994.tb00469.x. [DOI] [PubMed] [Google Scholar]
  11. Jucker B. A., Harms H., Zehnder A. J. Adhesion of the positively charged bacterium Stenotrophomonas (Xanthomonas) maltophilia 70401 to glass and Teflon. J Bacteriol. 1996 Sep;178(18):5472–5479. doi: 10.1128/jb.178.18.5472-5479.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Keen N. T., Tamaki S., Kobayashi D., Trollinger D. Improved broad-host-range plasmids for DNA cloning in gram-negative bacteria. Gene. 1988 Oct 15;70(1):191–197. doi: 10.1016/0378-1119(88)90117-5. [DOI] [PubMed] [Google Scholar]
  13. Knutton S., Lloyd D. R., McNeish A. S. Adhesion of enteropathogenic Escherichia coli to human intestinal enterocytes and cultured human intestinal mucosa. Infect Immun. 1987 Jan;55(1):69–77. doi: 10.1128/iai.55.1.69-77.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Knutton S., Shaw R. K., Bhan M. K., Smith H. R., McConnell M. M., Cheasty T., Williams P. H., Baldwin T. J. Ability of enteroaggregative Escherichia coli strains to adhere in vitro to human intestinal mucosa. Infect Immun. 1992 May;60(5):2083–2091. doi: 10.1128/iai.60.5.2083-2091.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kuehn M. J., Ogg D. J., Kihlberg J., Slonim L. N., Flemmer K., Bergfors T., Hultgren S. J. Structural basis of pilus subunit recognition by the PapD chaperone. Science. 1993 Nov 19;262(5137):1234–1241. doi: 10.1126/science.7901913. [DOI] [PubMed] [Google Scholar]
  16. Le Bouguenec C., Garcia M. I., Ouin V., Desperrier J. M., Gounon P., Labigne A. Characterization of plasmid-borne afa-3 gene clusters encoding afimbrial adhesins expressed by Escherichia coli strains associated with intestinal or urinary tract infections. Infect Immun. 1993 Dec;61(12):5106–5114. doi: 10.1128/iai.61.12.5106-5114.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Levine M. M., Bergquist E. J., Nalin D. R., Waterman D. H., Hornick R. B., Young C. R., Sotman S. Escherichia coli strains that cause diarrhoea but do not produce heat-labile or heat-stable enterotoxins and are non-invasive. Lancet. 1978 May 27;1(8074):1119–1122. doi: 10.1016/s0140-6736(78)90299-4. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Lukomski S., Hull R. A., Hull S. I. Identification of the O antigen polymerase (rfc) gene in Escherichia coli O4 by insertional mutagenesis using a nonpolar chloramphenicol resistance cassette. J Bacteriol. 1996 Jan;178(1):240–247. doi: 10.1128/jb.178.1.240-247.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Manoil C., Beckwith J. TnphoA: a transposon probe for protein export signals. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8129–8133. doi: 10.1073/pnas.82.23.8129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem. 1987 Jul 25;262(21):10035–10038. [PubMed] [Google Scholar]
  22. Nataro J. P., Baldini M. M., Kaper J. B., Black R. E., Bravo N., Levine M. M. Detection of an adherence factor of enteropathogenic Escherichia coli with a DNA probe. J Infect Dis. 1985 Sep;152(3):560–565. doi: 10.1093/infdis/152.3.560. [DOI] [PubMed] [Google Scholar]
  23. Nataro J. P., Deng Y., Cookson S., Cravioto A., Savarino S. J., Guers L. D., Levine M. M., Tacket C. O. Heterogeneity of enteroaggregative Escherichia coli virulence demonstrated in volunteers. J Infect Dis. 1995 Feb;171(2):465–468. doi: 10.1093/infdis/171.2.465. [DOI] [PubMed] [Google Scholar]
  24. Nataro J. P., Deng Y., Maneval D. R., German A. L., Martin W. C., Levine M. M. Aggregative adherence fimbriae I of enteroaggregative Escherichia coli mediate adherence to HEp-2 cells and hemagglutination of human erythrocytes. Infect Immun. 1992 Jun;60(6):2297–2304. doi: 10.1128/iai.60.6.2297-2304.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nataro J. P., Hicks S., Phillips A. D., Vial P. A., Sears C. L. T84 cells in culture as a model for enteroaggregative Escherichia coli pathogenesis. Infect Immun. 1996 Nov;64(11):4761–4768. doi: 10.1128/iai.64.11.4761-4768.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nataro J. P., Kaper J. B., Robins-Browne R., Prado V., Vial P., Levine M. M. Patterns of adherence of diarrheagenic Escherichia coli to HEp-2 cells. Pediatr Infect Dis J. 1987 Sep;6(9):829–831. doi: 10.1097/00006454-198709000-00008. [DOI] [PubMed] [Google Scholar]
  27. Nataro J. P., Scaletsky I. C., Kaper J. B., Levine M. M., Trabulsi L. R. Plasmid-mediated factors conferring diffuse and localized adherence of enteropathogenic Escherichia coli. Infect Immun. 1985 May;48(2):378–383. doi: 10.1128/iai.48.2.378-383.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nataro J. P., Yikang D., Giron J. A., Savarino S. J., Kothary M. H., Hall R. Aggregative adherence fimbria I expression in enteroaggregative Escherichia coli requires two unlinked plasmid regions. Infect Immun. 1993 Mar;61(3):1126–1131. doi: 10.1128/iai.61.3.1126-1131.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Nataro J. P., Yikang D., Yingkang D., Walker K. AggR, a transcriptional activator of aggregative adherence fimbria I expression in enteroaggregative Escherichia coli. J Bacteriol. 1994 Aug;176(15):4691–4699. doi: 10.1128/jb.176.15.4691-4699.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nowicki B., Labigne A., Moseley S., Hull R., Hull S., Moulds J. The Dr hemagglutinin, afimbrial adhesins AFA-I and AFA-III, and F1845 fimbriae of uropathogenic and diarrhea-associated Escherichia coli belong to a family of hemagglutinins with Dr receptor recognition. Infect Immun. 1990 Jan;58(1):279–281. doi: 10.1128/iai.58.1.279-281.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Prentki P., Krisch H. M. In vitro insertional mutagenesis with a selectable DNA fragment. Gene. 1984 Sep;29(3):303–313. doi: 10.1016/0378-1119(84)90059-3. [DOI] [PubMed] [Google Scholar]
  32. Savarino S. J., Fasano A., Robertson D. C., Levine M. M. Enteroaggregative Escherichia coli elaborate a heat-stable enterotoxin demonstrable in an in vitro rabbit intestinal model. J Clin Invest. 1991 Apr;87(4):1450–1455. doi: 10.1172/JCI115151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Savarino S. J., Fox P., Deng Y., Nataro J. P. Identification and characterization of a gene cluster mediating enteroaggregative Escherichia coli aggregative adherence fimbria I biogenesis. J Bacteriol. 1994 Aug;176(16):4949–4957. doi: 10.1128/jb.176.16.4949-4957.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Smith H. R., Scotland S. M., Willshaw G. A., Rowe B., Cravioto A., Eslava C. Isolates of Escherichia coli O44:H18 of diverse origin are enteroaggregative. J Infect Dis. 1994 Dec;170(6):1610–1613. doi: 10.1093/infdis/170.6.1610. [DOI] [PubMed] [Google Scholar]
  35. Taylor R. K., Manoil C., Mekalanos J. J. Broad-host-range vectors for delivery of TnphoA: use in genetic analysis of secreted virulence determinants of Vibrio cholerae. J Bacteriol. 1989 Apr;171(4):1870–1878. doi: 10.1128/jb.171.4.1870-1878.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Tzipori S., Montanaro J., Robins-Browne R. M., Vial P., Gibson R., Levine M. M. Studies with enteroaggregative Escherichia coli in the gnotobiotic piglet gastroenteritis model. Infect Immun. 1992 Dec;60(12):5302–5306. doi: 10.1128/iai.60.12.5302-5306.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Vial P. A., Mathewson J. J., DuPont H. L., Guers L., Levine M. M. Comparison of two assay methods for patterns of adherence to HEp-2 cells of Escherichia coli from patients with diarrhea. J Clin Microbiol. 1990 May;28(5):882–885. doi: 10.1128/jcm.28.5.882-885.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Vial P. A., Robins-Browne R., Lior H., Prado V., Kaper J. B., Nataro J. P., Maneval D., Elsayed A., Levine M. M. Characterization of enteroadherent-aggregative Escherichia coli, a putative agent of diarrheal disease. J Infect Dis. 1988 Jul;158(1):70–79. doi: 10.1093/infdis/158.1.70. [DOI] [PubMed] [Google Scholar]
  39. Wanke C. A., Schorling J. B., Barrett L. J., Desouza M. A., Guerrant R. L. Potential role of adherence traits of Escherichia coli in persistent diarrhea in an urban Brazilian slum. Pediatr Infect Dis J. 1991 Oct;10(10):746–751. doi: 10.1097/00006454-199110000-00006. [DOI] [PubMed] [Google Scholar]
  40. Yamamoto T., Endo S., Yokota T., Echeverria P. Characteristics of adherence of enteroaggregative Escherichia coli to human and animal mucosa. Infect Immun. 1991 Oct;59(10):3722–3739. doi: 10.1128/iai.59.10.3722-3739.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES