The Locus for Enterocyte Effacement (Lee) of Enteropathogenic Escherichia Coli (Epec) from Dogs and Cats

  • Frédéric Goffaux
  • Bernard China
  • Laurence Janssen
  • Vinciane Pirson
  • Jacques Mainil
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 473)


Enteropathogenic Escherichia coli (EPEC) produce attaching and effacing lesions. The genes responsible for this lesion are clustered on the chromosome forming a 35.5 kilobase pathogenesis island called LEE. The LEE was identified, characterized and completely sequenced from the human EPEC strain E2348/69. The LEE carries genes coding for: a type III secretion system (genes esc and sep), the translocated intimin receptor (gene tir), the outer membrane protein intimin (gene eae) and the E. coli secreted proteins EspA, EspB, and EspD (genes esp). In addition to man and farm animals, EPEC are also isolated from dogs and cats. We studied structurally and functionally the LEE of dog and cat EPEC. First, we used four probes scattered along the LEE to identify the presence of a LEE in canine and feline EPEC isolates. Second, by PCR, we checked the presence of genes homologous to eae, sep, esp, and tir genes in these strains. Third, since the four types of eae and tir genes were described, we developed a multiplex PCR in order to determine the type of eae and tir genes present in each strain. Fourth, we determined by PCR the site of the LEE insertion on the chromosome. Fifth, we tested several of the canine-EPEC in their capacity to induce attaching and effacing lesions in the rabbit intestinal loop assay. We can conclude from this study: first, that the a LEE-like structure is present in all tested strains and that it contains genes homologous to esp, sep, tir, and eae genes; second, that there is some preferential associations between the type of eae gene and the type of tir gene present in a strain; third, that the majority of the tested strains contained a LEE located elsewhere on the chromosome in comparison to the human EPEC strain E2348/69; and fourth that dog EPEC were able to induce attaching and effacing lesions in rabbit ileal loop assay.


Pathogenicity Island Enterocyte Effacement Intimate Attachment Loop Assay Epithelial Brush Border 
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  1. Abe, A., Kenny, B., Stein, M., and Finlay, B.B., 1997, Characterization of two virulence proteins secreted by rabbit enteropathogenic Escherichia coli, EspA and EspB, whose maximal expression is sensitive to host body temperature, Infect. Immun., 65:3547–3555.PubMedGoogle Scholar
  2. Agin, T.S., Cantey, J.R., Boeedeker, E.C., and Wolf, M.K., 1996, Characterization of the eaeA gene from rabbit enteropathogenic Escherichia coli RDEC-1 and comparison to other eaeA genes from bacteria that cause attaching-afacing lesions, FEMS Microbiol. Lett., 14:249–258.CrossRefGoogle Scholar
  3. An, H., Fairbrother, J.M., Dubreuil, D., and Harel, J., 1997, Cloning and characterization of the eae gene from a dog attaching and effacing Escherichia coli strain 4221, FEMS Microbiol. Lett., 148:239–245.PubMedCrossRefGoogle Scholar
  4. Beebakhee, G., Louie, M., De Azavedo, J., and Brunton, J., 1992, Cloning and nucleotide sequence of the eae gene homologue from enterohemorrhagic Escherichia coli serotype O157:H7. FEMS Microbiol. Lett., 91:63–68.CrossRefGoogle Scholar
  5. Broes, A., Drolet, R., Fairbrother, J.M., and Johnson, W.M., 1988, Natural infection with attaching and effacing Escherichia coli in a diarrheic puppy, Can. J. Vet. Res., 52:280–282.PubMedGoogle Scholar
  6. China, B., Pirson, V., and Mainil, J., 1996, Typing of bovine attaching and effacing Escherichia coli by multiplex in vitro amplification of virulence-associated genes, Appl. Environ. Microbiol., 62:3462–3465.PubMedGoogle Scholar
  7. China, B., Pirson, V., Jacquemin, E., Pohl, P., and Mainil, J.G., 1997, Pathotypes of bovine verotoxigenic Escherichia coli isolates producing attaching/Effacing (AE) lesions in the ligated inbtestinal loop assay in rabbits, In Mechanisms in the pathogenesis of enteric diseases, Editors: Paul, P.S., Francis, D.H., and Benfield, D.A., Adv. Exp. Med. Biol. 412, pp. 311–316.Google Scholar
  8. Donnenberg, M.S., Yu, J., and Kaper, J.B., 1993, A second chromosomal gene necessary for intimate attachment of enteropahtogenic Escherichia coli to epithelial cells, J Bacteriol., 175:4670–4680.PubMedGoogle Scholar
  9. Drolet, R., Fairbrother, J.M., Harel, J., and Helie, P., 1994, Attaching and effacing and enterotoxigenic Escherichia coli associated with enteric colibacillosis in the dog, Can. J. Vet. Res., 31:591–594.Google Scholar
  10. Elliott, S.X, Wainwright, L.A., McDaniel, T.K., Jarvis, K.G., Deng, Y.K., Lai, L.C., McNamara, B.P., Donnenberg, M.S., and Kaper, J.B., 1998, The complete sequence of the locus of enterocyte effacement (LEE) from enteropathogenic Escherichia coli E2348/69, Mol. Microbiol., 28:1–4.PubMedCrossRefGoogle Scholar
  11. Finlay, B.B., Rosenshine, I., Donnenberg, M.S., and Kaper, XB., 1992, Cytoskeletal composition of attaching and effacing lesions associated with enteropatogenic Escherichia coli adherence to HeLa cells, Infect. Immun., 60:2541–2543.PubMedGoogle Scholar
  12. Goffaux, F., Mainil, J., Pirson, V., Charlier, G., Pohl, P., Jacquemin, E., and China, B., 1997, Bovine attaching and effacing Escherichia coli possess a pathogenesis island related to the LEE of the human enteropathogenic E. coli strain E2348/69, FEMS Microbiol. Lett., 154:415–421.PubMedCrossRefGoogle Scholar
  13. Janke, B.H., Francis, D.H., Collins, J.E., Libal, M.C., Zeman, D.H., and Johnson, D.D., 1989, Attaching and effacing Escherichia coli infections in calves, pigs, lambs and dogs, J. Vet. Diagn. Invest., 1:6–11.PubMedCrossRefGoogle Scholar
  14. Jarvis, K.G., Giron, J.A., Jerse, A.E., McDanie, T.K., Donnenber, M.S., and Kaper, J.B., 1995, Enteropathogenic Escherichia coli contains a putative type III secretion system necessary for the export of proteins involved in attaching and effacing lesion formation. Proc. Natl. Acad. Sci. USA, 92:7996–8000.PubMedCrossRefGoogle Scholar
  15. Jerse, A.E., Yu, J., Tall, B.D., and Kaper, J.B., 1990, A genetic locus of enteropathogenic Escherichia coli necessary for the production of ataching and effacing lesions on tissue culture cells. Proc. Natl. Acad. Sci. USA, 87:7839–7843.PubMedCrossRefGoogle Scholar
  16. Kenny, B., Lai, L.-C., Finlay, B.B., and Donnenberg, M.S., 1996, EspA, a protein secreted by enteropathogenic Escherichia coli is required to induces in epithelial epithelial cells. Mol. Microbiol., 20:313–323.PubMedCrossRefGoogle Scholar
  17. Kenny, B., DeVinney, R., Stein, M., Reinscheid, D.X, Frey, E.A., and Finlay, B.B., 1997, Enteropathogenic E. coli (EPEC) transfers its receptor for intimate adherence into mammalian cells, Cell, 91:511–520.PubMedCrossRefGoogle Scholar
  18. Knutton, S., Rosenshine, I., Pallen, M.J., Nisan, I., Neves, B.C., Bain, G, Wolff, G, Dougan, G., and Frankel, G., 1998, A novel EspA-associated surface organelle of enteropathogenic Escherichia coli; involved in protein translocation into epithelial cells. EMBO J., 17:2166–2176.PubMedCrossRefGoogle Scholar
  19. Knutton, S., Attaching and Effacing E. Coli. In: Escherichia coli in domestic animals and humans, Editor: Gyles, C.L., 1994, CAB International, Wallingford, UK, pp. 567–591.Google Scholar
  20. Lai, L.C., Wainwright, L.A., Stone, K.D., and Donnenberg, M.S., 1997, A third secreted protein that is encoded by the enteropazthogenic Escherichia coli pathogenicity island is required for transduction of signals and for attaching and effacing activities in host cells. Infect. Immun., 65:2211–2217.PubMedGoogle Scholar
  21. Mainil, J.G., Jacquemin, E., Bez, S., Pohl, P., and Kaeckenbeeck, A., 1994, La détection, au moyen de sondes génétiques, de souches potentiellement attachantes et effaçantes & Escherichia coli (AEEC) isolées de veaux, de porcelets et de carnivores. In: biotechnologies du diagnostic et de la prévention des maladies animales, Editor: AUPELF-UREF, John Libbey Eurotext, Paris, France, pp. 73–79.Google Scholar
  22. McDaniel, T.K., Jarvis, G, Donnenberg, M.S., and Kaper, J.B., 1995, A genetic locus of enterocyte effacement conserved among diverse enterobacterial pathogens. Proc. Natl. Acad. Sci. USA, 92:1664–1668.PubMedCrossRefGoogle Scholar
  23. McDaniel, T.K. and Kaper, J.B., 1997, A cloned pathogenicity island from enteropathogenic Escherichia coli confers the attaching and effacing phenotype on E. coli K-12, Mol. Microbiol., 23:399–407.PubMedCrossRefGoogle Scholar
  24. Moon, H.W., Whipp, S.C., Argenzio, R.A., Levine, M.M., and Gianella, R.A., 1983, Attaching and effacing activities of rabbit and human enteropathogenic Escherichia coli in pig and rabbit intestines. Infect. Immun., 53:1340–1351.Google Scholar
  25. Nataro, J.P. and Kaper, J.B., 1998, Diarrheagic Escherichia coli. Clin. Microbiol. Rev., 11:142–201.PubMedGoogle Scholar
  26. Paton, A.W., Manning, P.A., Woodrow, M.C., and Paton J.C., 1998, The translocated intimin receptor (Tir) of shiga toxigenic Escherichia coli isolates belonging to serotype O26, Olli, and O157 react with sera from patients with hemolytic-uremic syndrome and exhibit marked sequence heterogeneity. Genbank acession number AF070067-AF070068-AF070069.Google Scholar
  27. Perna, N., Mayhew, G.F., Posfai, G, Elliot, S., Donnenberg, M.S., Kaper, J.B., and Blattner, ER., 1998, Molecular evolution of a pathogenicity island from enterohemorrhagic Escherichia coli O157:H7. Infect. Immun., 66:3810–3817.PubMedGoogle Scholar
  28. Popischil, A., Mainil, J.G., Baljer, G, and Moon, H.W., 1987, Attaching and effacing bacteria in the intestines of calves and cats with diarrhea, Vet. Path., 24:330–334.Google Scholar
  29. Yu, J. and Kaper, J.B., 1992, Cloning and characterization of the eae gene of enterohaemorrhagic Escherichia coli O157:H7. Mol. Microbiol., 6:411–471.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1999

Authors and Affiliations

  • Frédéric Goffaux
    • 1
  • Bernard China
    • 1
  • Laurence Janssen
    • 1
  • Vinciane Pirson
    • 1
  • Jacques Mainil
    • 1
  1. 1.Laboratory of Bacteriology Faculty of Veterinary MedicineUniversity of LiègeLiègeBelgium

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