Subtractive hybridization and identification of putative adhesins in a Shiga toxin-producing eae-negative Escherichia coli

Maricel Vidal, Valeria Prado, Gregory C. Whitlock, Aldo Solari, Alfredo Torres, Roberto M. Vidal

Research output: Contribution to journalArticle

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Abstract

Adherence to epithelial cells by specific adhesins is a characteristic of Shiga toxin-producing Escherichia coli (STEC) strains. The eae-encoded protein intimin is the main adhesin implicated in intestinal colonization in vivo.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 eae-negative and still adhere to epithelial cells at a level 100-fold higher than enterohaemorrhagic E. coli (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 eae-negative strain 472-1 was used to identify regions implicated in adhesion. In addition to the saa gene, we identified 18 specific genes in STEC 472-1, 16 of which had nucleotide identity to Salmonella ST46 phage genes; the two remaining ones shared identity to a gene encoding a hypothetical protein of uropathogenic E. coli. The DNA sequence of the STEC 472-1 psu-int region identified five open reading frames with homology to phage genes. We constructed mutant strains in the saa gene and the psu-int region to study the participation of these genes in the adherence to epithelial cells and our results demonstrated that STECΔsaa and STECΔpsu-int 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 eae-independent matter and that saa and psu-int participate in this adhesion process.

Original languageEnglish
Pages (from-to)3639-3648
Number of pages10
JournalMicrobiology
Volume154
Issue number12
DOIs
StatePublished - 2008

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Shiga Toxin
Shiga-Toxigenic Escherichia coli
Escherichia coli
Genes
Epithelial Cells
Enterohemorrhagic Escherichia coli
Salmonella Phages
Uropathogenic Escherichia coli
Proteins
Escherichia coli O157
Subtractive Hybridization Techniques
Chile
Bacteriophages
Open Reading Frames
Nucleotides
Phenotype

ASJC Scopus subject areas

  • Microbiology

Cite this

Subtractive hybridization and identification of putative adhesins in a Shiga toxin-producing eae-negative Escherichia coli. / Vidal, Maricel; Prado, Valeria; Whitlock, Gregory C.; Solari, Aldo; Torres, Alfredo; Vidal, Roberto M.

In: Microbiology, Vol. 154, No. 12, 2008, p. 3639-3648.

Research output: Contribution to journalArticle

Vidal, Maricel ; Prado, Valeria ; Whitlock, Gregory C. ; Solari, Aldo ; Torres, Alfredo ; Vidal, Roberto M. / Subtractive hybridization and identification of putative adhesins in a Shiga toxin-producing eae-negative Escherichia coli. In: Microbiology. 2008 ; Vol. 154, No. 12. pp. 3639-3648.
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