Enteropathogenic Escherichia coli infection leads to appearance of aberrant tight junctions strands in the lateral membrane of intestinal epithelial cells

Cell Microbiol. 2004 Aug;6(8):783-93. doi: 10.1111/j.1462-5822.2004.00404.x.


Infection of intestinal epithelial cells with enteropathogenic Escherichia coli (EPEC) disrupts tight junction (TJ) architecture and barrier function. The aim of this study was to determine the impact of EPEC on TJ protein interactions and localization. Human intestinal epithelial cells (T84) were infected for 1, 3 or 6 h with EPEC. To probe the TJ protein-protein interactions, co-immunoprecipitations were performed. The associations between ZO-1, occludin and claudin-1 progressively decreased after infection. Corresponding morphological changes were analysed by immunofluorescence confocal microscopy. Tight junction proteins progressively lost their apically restricted localization. Freeze-fracture electron microscopy revealed the appearance of aberrant strands throughout the lateral membrane that contained claudin-1 and occludin as determined by immunogold labelling. These structural alterations were accompanied by a loss of barrier function. Mutation of the gene encoding EspF, important in the disruption of TJs by EPEC, prevented the disruption of TJs. Tight junction structure normalized following eradication of EPEC with gentamicin and overnight recovery. This is the first demonstration that a microbial pathogen can cause aberrant TJ strands in the lateral membrane of host cells. We speculate that the disruption of integral and cytoplasmic TJ protein interactions following EPEC infection allows TJ strands to form or diffuse into the lateral plasma membrane.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cell Line
  • Cell Membrane / metabolism
  • Cell Membrane / microbiology
  • Cell Membrane / ultrastructure
  • Claudin-1
  • Epithelial Cells / metabolism
  • Epithelial Cells / microbiology*
  • Epithelial Cells / ultrastructure
  • Escherichia coli / pathogenicity*
  • Freeze Fracturing
  • Humans
  • Intestine, Small / metabolism
  • Intestine, Small / microbiology*
  • Intestine, Small / ultrastructure
  • Membrane Proteins / metabolism
  • Microscopy, Electron
  • Occludin
  • Phosphoproteins / metabolism
  • Protein Interaction Mapping
  • Tight Junctions / microbiology*
  • Tight Junctions / ultrastructure
  • Zonula Occludens-1 Protein


  • CLDN1 protein, human
  • Claudin-1
  • Membrane Proteins
  • OCLN protein, human
  • Occludin
  • Phosphoproteins
  • TJP1 protein, human
  • Zonula Occludens-1 Protein