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Volume 154, Issue 12

Subtractive hybridization and identification of putative adhesins in a Shiga toxin-producing -negative Free

Abstract

Adherence to epithelial cells by specific adhesins is a characteristic of Shiga toxin-producing (STEC) strains. The -encoded protein intimin is the main adhesin implicated in intestinal colonization . We recently showed that STEC strains isolated in Chile displayed a wide variety of adhesins; here we demonstrate that some of these STEC strains are -negative and still adhere to epithelial cells at a level 100-fold higher than enterohaemorrhagic (EHEC) O157 : H7 prototype strain EDL933. This phenotype is associated with the presence of adherence factors different from the intimin protein. Subtractive hybridization between EHEC EDL933 and STEC -negative strain 472-1 was used to identify regions implicated in adhesion. In addition to the gene, we identified 18 specific genes in STEC 472-1, 16 of which had nucleotide identity to ST46 phage genes; the two remaining ones shared identity to a gene encoding a hypothetical protein of uropathogenic . The DNA sequence of the STEC 472-1 region identified five open reading frames with homology to phage genes. We constructed mutant strains in the gene and the region to study the participation of these genes in the adherence to epithelial cells and our results demonstrated that STECΔ and STECΔ mutants displayed a 10-fold decrease in adherence as compared to the STEC 472-1 wild-type strain. Overall, our results suggest that STEC strain 472-1 adheres to epithelial cells in an -independent matter and that and participate in this adhesion process.

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Keyword(s): A/E, attaching and effacing , EHEC, enterohaemorrhagic E. coli , HC, haemorrhagic colitis , HUS, haemolytic uraemic syndrome , LEE, locus of enterocyte effacement and STEC, Shiga toxin-producing Escherichia coli
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2008-12-01
2024-04-20
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References

  1. Bertin Y., Boukhors K., Livrelli V., Martin Ch. 2004; Localization of the insertion site and pathotype determination of the locus of enterocyte effacement of Shiga toxin-producing Escherichia coli strains. Appl Environ Microbiol 70:61–68
     [Google Scholar]
  2. Beutin L., Geier D., Steinrück H., Zimmerman S., Scheutz F. J. 1993; Prevalence and some properties of verotoxin (Shiga-like toxin)-producing Escherichia coli in seven different species of healthy domestic animals. J Clin Microbiol 31:2483–2488
     [Google Scholar]
  3. Blanco J., Blanco M., Blanco J. E., Mora A., González E., Bernárdez M., Alonso M., Coira A., Rodríguez A. other authors 2003; Verotoxin-producing Escherichia coli in Spain: prevalence, serotypes, and virulence genes of O157 : H7 and non-O157 VTEC in ruminants, raw beef products, and humans. Exp Biol Med (Maywood 228:345–351
     [Google Scholar]
  4. Blanco M., Schumacher S., Tasara T., Zweifel C., Blanco J. E., Dahbi G., Blanco J., Stephan R. 2005; Serotypes, intimin variants and other virulence factors of eae positive Escherichia coli strains isolated from healthy cattle in Switzerland. Identification of a new intimin variant gene ( eae- 2). BMC Microbiol 5:23–33
     [Google Scholar]
  5. Boerlin P., McEwen S., Boerlin-Petzold F., Wilson J., Johnson R., Gyles C. 1999; Associations between virulence factors of Shiga toxin-producing Escherichia coli and disease in humans. J Clin Microbiol 37:497–503
     [Google Scholar]
  6. Brunder W., Khan A. S., Hacker J., Karch H. 2001; Novel type of fimbriae encoded by the large plasmid of sorbitol-fermenting enterohemorrhagic Escherichia coli O157 : H–. Infect Immun 69:4447–4457
     [Google Scholar]
  7. Calia K. E., Waldor M. K., Calderwood S. B. 1998; Use of representational difference analysis to identify genomic differences between pathogenic strains of Vibrio cholerae . Infect Immun 66:849–852
     [Google Scholar]
  8. Caprioli A., Nigrelli A., Gatti R., Zavanella M., Blando A. M., Minelli F., Donelli G. 1993; Characterisation of verocytotoxin-producing Escherichia coli isolated from pigs and cattle in northern Italy. Vet Rec 133:323–324
     [Google Scholar]
  9. Caprioli A., Morabito S., Brugère H., Oswald E. 2005; Enterohaemorrhagic Escherichia coli: emerging issues on virulence and modes of transmission. Vet Res 36:289–311
     [Google Scholar]
  10. Datsenko K. A., Wanner B. L. 2000; One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci U S A 97:6640–6645
     [Google Scholar]
  11. De Azavedo J., McWhirter E., Louie M., Brunton J. 1994; Eae-negative verotoxin-producing Escherichia coli associated with haemolytic uremic syndrome and hemorrhagic colitis. In Recent Advances in Verocytotoxin-Producing Escherichia coli Infections pp 265–268 Edited by Karmali M. A., Goglio A. G. Amsterdam, The Netherlands: Elsevier Science B.V;
     [Google Scholar]
  12. Donnenberg M. S., Tacket C. O., James S. P., Losonky G., Nataro J. P., Wasserman S. S., Kaper J. B., Levine M. M. 1993; Role of the eaeA gene in experimental enteropathogenic Escherichia coli infection. J Clin Invest 92:1412–1417
     [Google Scholar]
  13. Dytoc M. T., Ismaili A., Philpott D. J., Soni R., Brunton J. L., Sherman P. M. 1994; Distinct binding properties of eae negative verocytotoxin-producing Escherichia coli of serotype O113 : H21. Infect Immun 62:3494–3505
     [Google Scholar]
  14. Heermann R., Fuchs T. M. 2008; Comparative analysis of the Photorhabdus luminescens and the Yersinia enterocolitica genomes: uncovering candidate genes involved in insect pathogenicity. BMC Genomics 9:40
     [Google Scholar]
  15. Kaper J. B., Gansheroff L. J., Wachtel M. R., O'Brien A. D. 1998; Intimin-mediated adherence of Shiga toxin-producing Escherichia coli and attaching-and-effacing pathogens. In Escherichia coli O157 : H7 and Other Shiga Toxin-Producing E. coli Strains pp 148–156 Edited by Kaper J. B., O'Brien A. D. Washington, DC: American Society for Microbiology;
     [Google Scholar]
  16. Karmali M. A. 1989; Infection by verocytotoxin-producing Escherichia coli . Clin Microbiol Rev 2:15–38
     [Google Scholar]
  17. Lucchesi P. M., Kruger A., Parma A. E. 2005; Distribution of saa gene variants in verocytotoxigenic Escherichia coli isolated from cattle and food. Res Microbiol 157:263–266
     [Google Scholar]
  18. Nicholls L., Grant T. H., Robins-Browne R. M. 2000; Identification of a novel genetic locus that is required for in vitro adhesion of a clinical isolate of enterohemorrhagic Escherichia coli to epithelial cells. Mol Microbiol 35:275–288
     [Google Scholar]
  19. Paton J. C., Paton A. W. 1998; Pathogenesis and diagnosis of Shiga toxin-producing Escherichia coli infections. Clin Microbiol Rev 11:450–479
     [Google Scholar]
  20. Paton A. W., Woodrow M. C., Doyle R. M., Lanser J. A., Paton J. C. 1999; Molecular characterization of a shiga toxigenic Escherichia coli O113 : H21 strain lacking eae responsible for a cluster of cases of hemolytic-uremic syndrome. J Clin Microbiol 37:3357–3361
     [Google Scholar]
  21. Paton A. W., Srimanote P., Woodrow M. C., Paton J. C. 2001; Characterization of Saa, a novel autoagglutinating adhesin produced by locus enterocyte effacement-negative shiga-toxigenic Escherichia coli strains that are virulent for humans. Infect Immun 69:6999–7009
     [Google Scholar]
  22. Pierson L. S. III, Kahn M. L. 1987; Integration of satellite bacteriophage P4 in Escherichia coli. DNA sequences of the phage and host regions involved in site-specific recombination. J Mol Biol 196:487–496
     [Google Scholar]
  23. Scotland S., Knutton S., Said B., Rowe B. 1994; Adherence to Caco-2 cells of Vero cytotoxin-producing strains of Escherichia coli belonging to serogroups other than O157. In Recent Advances in Verocytotoxin-Producing Escherichia coli Infections pp 257–260 Edited by Karmali M. A., Goglio A. G. Amsterdam, The Netherlands: Elsevier Science B.V;
     [Google Scholar]
  24. Tarr P. I., Bilge S. S., Vary J. C. Jr, Jelacic S., Habeeb R. L., Ward T. R., Baylor M. R., Besser T. E. 2000; Iha: a novel Escherichia coli O157 : H7 adherence-conferring molecule encoded on a recently acquired chromosomal island of conserved structure. Infect Immun 68:1400–1407
     [Google Scholar]
  25. Thomson N., Baker S., Pickard D., Fookes M., Anjum M., Hamlin N., Wain J., House D., Bhutta Z. other authors 2004; The role of prophage-like elements in the diversity of Salmonella enterica serovars. J Mol Biol 339:279–300
     [Google Scholar]
  26. Toma C., Martínez E., Song T., Miliwebsky E., Chinen I., Iyoda S., Iwanaga M., Rivas M. 2004; Distribution of putative adhesin in different seropathotypes of Shiga toxin-producing Escherichia coli . J Clin Microbiol 42:4937–4946
     [Google Scholar]
  27. Torres A. G., Girón J., Perna N., Burland V., Blattner F., Avelino-Flores F., Kaper J. 2002; Identification and characterization of lpfABCC′DE, a fimbrial operon of enterohemorrhagic Escherichia coli O157 : H7. Infect Immun 70:5416–5427
     [Google Scholar]
  28. Vidal M., Escobar P., Prado V., Hormazabal J. C., Vidal R. 2007; Distribution of putative adhesins in Shiga toxin-producing Escherichia coli (STEC) strains isolated from different sources in Chile. Epidemiol Infect 135:688–694
     [Google Scholar]
  29. Verweyen H. M., Karch H., Brandis M., Zimmerhackl L. B. 2000; Enterohemorrhagic Escherichia coli infections: following transmission routes. Pediatr Nephrol 14:73–83
     [Google Scholar]
  30. Zhang Y. L., Ong C. T., Leung K. Y. 2000; Molecular analysis of genetic differences between virulent and avirulent strains of Aeromonas hydrophila isolated from diseased fish. Microbiology 146:999–1009
     [Google Scholar]
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Subtractive hybridization and identification of putative adhesins in a Shiga toxin-producing eae-negative Escherichia coli
Microbiology 154, 3639 (2008); https://doi.org/10.1099/mic.0.2008/021212-0
/content/journal/micro/10.1099/mic.0.2008/021212-0
/content/journal/micro/10.1099/mic.0.2008/021212-0
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