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PMID: 14617638 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Translocated intimin receptor and its chaperone interact with ATPase of the type III secretion apparatus of enteropathogenic Escherichia coli.

Journal of bacteriology ·Vol. 185 ·No. 23 ·2003-12-00 ·Pages 6747-55

Gauthier A, Finlay BB

Abstract

Few interactions have been reported between effectors and components of the type III secretion apparatus, although many interactions have been demonstrated between type III effectors and their cognate chaperones. It is thought that chaperones may play a role in directing effectors to the type III secretion apparatus. The ATPase FliI in the flagellar assembly apparatus plays a pivotal role in interacting with other components of the apparatus and with substrates of the flagellar system. We performed experiments to determine if there were any interactions between the effector Tir and its chaperone CesT and the type III secretion apparatus of enteropathogenic Escherichia coli (EPEC). Specifically, based on analogies with the flagella system, we examined Tir-CesT interactions with the putative ATPase EscN. We showed by affinity chromatography that EscN and Tir bind CesT specifically. Tir is not necessary for CesT and EscN interactions, and EscN binds Tir specifically without its chaperone CesT. Moreover, Tir directly binds EscN, as shown via gel overlay and enzyme-linked immunosorbent assay, and coimmunoprecipitation experiments revealed that Tir interacts with EscN inside EPEC. These data provide evidence for direct interactions between a chaperone, effector, and type III component in the pathogenic type III secretion system and suggest a model for Tir translocation whereby its chaperone, CesT, brings Tir to the type III secretion apparatus by specifically interacting with the type III ATPase EscN.

MeSH Terms
Adenosine Triphosphatases/metabolism Escherichia coli/enzymology,metabolism,pathogenicity Escherichia coli Proteins/metabolism Molecular Chaperones/metabolism Protein Binding Protein Transport Receptors, Cell Surface/metabolism Species Specificity
Chemicals
CesT protein, E coli Escherichia coli Proteins Molecular Chaperones Receptors, Cell Surface Tir protein, E coli Adenosine Triphosphatases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Gauthier Annick
Biotechnology Laboratory and Department of Biochemistry, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada.
Finlay B Brett
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/JB.185.23.6747-6755.2003
Published
2003-12-00
Pages
6747-55
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC262708
Subset
IM
External Links
PubMed source DOI: 10.1128/JB.185.23.6747-6755.2003 PMC Full Text View journal details
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