title: Enteropathogenic Escherichia coli (EPEC) Tir receptor molecule does not undergo full modification when introduced into host cells by EPEC-independent mechanisms.
abstract: Enteropathogenic Escherichia coli (EPEC), like many other gram-negative pathogens, encodes a type III secretion apparatus dedicated to the release of virulence-associated proteins. One such protein, Tir, is translocated into host cells, where it is modified by the addition of phosphate groups, resulting in a number of species with distinct molecular mass. One phosphorylation event, on tyrosine residue 474 of Tir, does not contribute to shifts in molecular mass but is essential for its actin-nucleating function. The role of the nonphosphotyrosine related modifications is unknown. In this paper, we demonstrate, using three different approaches, that Tir does not encode sufficient information to facilitate its complete modification when introduced into host cells in EPEC-independent mechanisms. Each system revealed that Tir is a substrate for a host kinase whose action results in its partial modification to a form similar to one evident in EPEC-infected host cells. Further Tir modification could not be induced by infecting cells with EPEC, suggesting that Tir must be coexpressed with other EPEC factors to enable its full modification within host cells. One approach used Yersinia spp. to deliver Tir into host cells, and this system revealed that Tir secretion and translocation can occur in the absence of the Tir chaperone molecule, CesT (formerly known as OrfU). CesT was found to be an efficiency factor which was not required, unlike in EPEC, for Tir stability, indicating that it may function to guide Tir to the translocation apparatus or maintain it in a secretion-competent form.
authors.affiliation: Department of Pathology and Microbiology, School of Medical Sciences, University Walk, Bristol BS8 1TD, England. B.Kenny@bristol.ac.uk
authors.forename: B
authors.initials: B
authors.lastname: Kenny
authors.forename: J
authors.initials: J
authors.lastname: Warawa
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D002463:Cell Membrane Permeability; D004926:Escherichia coli / Q000472:pathogenicity*; D029968:Escherichia coli Proteins; D006367:HeLa Cells; D006801:Humans; D007410:Intestinal Diseases / Q000382:microbiology; D018832:Molecular Chaperones / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D011956:Receptors, Cell Surface / Q000378:metabolism*; D015011:Yersinia pseudotuberculosis / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; C122044:CesT protein, E coli; D029968:Escherichia coli Proteins; D018832:Molecular Chaperones; D011956:Receptors, Cell Surface; C109287:Tir protein, E coli