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

Microbiology (Reading). 2008 Dec;154(Pt 12):3639-3648. doi: 10.1099/mic.0.2008/021212-0.

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 STECDeltasaa and STECDeltapsu-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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adhesins, Bacterial / genetics
  • Adhesins, Bacterial / metabolism*
  • Adhesins, Escherichia coli / chemistry
  • Adhesins, Escherichia coli / genetics
  • Adhesins, Escherichia coli / metabolism*
  • Bacterial Adhesion
  • Capsid Proteins / genetics
  • Capsid Proteins / metabolism
  • Cell Line
  • Epithelial Cells / microbiology
  • Escherichia coli O157 / genetics
  • Escherichia coli O157 / metabolism
  • Escherichia coli O157 / physiology
  • Escherichia coli Proteins / chemistry
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism*
  • Humans
  • Mutation
  • Nucleic Acid Hybridization / methods*
  • Shiga-Toxigenic Escherichia coli / genetics
  • Shiga-Toxigenic Escherichia coli / metabolism
  • Shiga-Toxigenic Escherichia coli / physiology*

Substances

  • Adhesins, Bacterial
  • Adhesins, Escherichia coli
  • Capsid Proteins
  • Escherichia coli Proteins
  • SAA protein, E coli
  • polarity suppression factor, bacteriophage P4
  • eaeA protein, E coli