Binding of intimin from enteropathogenic Escherichia coli to Tir and to host cells

Enteropathogenic Escherichia coli (EPEC) induce characteristic attaching and effacing (A/E) lesions on epithelial cells. This event is mediated, in part, by binding of the bacterial outer membrane protein, intimin, to a second EPEC protein, Tir (translocated intimin receptor), which is exported by t...

Full description

Saved in:
Bibliographic Details
Published in:Molecular microbiology 1999-04, Vol.32 (1), p.151-158
Main Authors: Hartland, Elizabeth L., Batchelor, Miranda, Delahay, Robin M., Hale, Christine, Matthews, Stephen, Dougan, Gordon, Knutton, Stuart, Connerton, Ian, Frankel, Gad
Format: Article
Language:English
Subjects:
Adhesins, Bacterial
Bacterial Adhesion
Bacterial Outer Membrane Proteins - metabolism
beta-Galactosidase - metabolism
Carrier Proteins
Cell Line
Cloning, Molecular
Escherichia coli
Escherichia coli - metabolism
Escherichia coli - pathogenicity
Escherichia coli Proteins
HeLa Cells
Humans
Immunohistochemistry
Microscopy, Electron
Microscopy, Fluorescence
Microscopy, Immunoelectron
Models, Genetic
Models, Molecular
Plasmids
Protein Binding
Receptors, Cell Surface - metabolism
Yeasts - genetics
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Enteropathogenic Escherichia coli (EPEC) induce characteristic attaching and effacing (A/E) lesions on epithelial cells. This event is mediated, in part, by binding of the bacterial outer membrane protein, intimin, to a second EPEC protein, Tir (translocated intimin receptor), which is exported by the bacteria and integrated into the host cell plasma membrane. In this study, we have localized the intimin‐binding domain of Tir to a central 107‐amino‐acid region, designated Tir‐M. We provide evidence that both the amino‐ and carboxy‐termini of Tir are located within the host cell. In addition, using immunogold labelling electron microscopy, we have confirmed that intimin can bind independently to host cells even in the absence of Tir. This Tir‐independent interaction and the ability of EPEC to induce A/E lesions requires an intact lectin‐like module residing at the carboxy‐terminus of the intimin polypeptide. Using the yeast two‐hybrid system and gel overlays, we show that intimin can bind both Tir and Tir‐M even when the lectin‐like domain is disrupted. These data provide strong evidence that intimin interacts not only with Tir but also in a lectin‐like manner with a host cell intimin receptor.
ISSN:0950-382X
1365-2958
DOI:10.1046/j.1365-2958.1999.01338.x