title: CesT is a bivalent enteropathogenic Escherichia coli chaperone required for translocation of both Tir and Map.
abstract: Map is an enteropathogenic Escherichia coli (EPEC) protein that is translocated into eukaryotic cells by a type III secretion system. Although not required for the induction of attaching and effacing (A/E) lesion formation characteristic of EPEC infection, translocated Map is suggested to disrupt mitochondrial membrane potential, which may impact upon subsequent functions of the organelle such as control of cell death. Before secretion, many effector proteins are maintained in the bacterial cytosol by association with a specific chaperone. In EPEC, chaperones have been identified for the effector proteins translocated intimin receptor (Tir) and EspF, and for the translocator proteins EspB and EspD. In this study, we present evidence that the Tir-specific chaperone, CesT, also performs a chaperone function for Map. Using a combination of biochemical approaches, we demonstrate specific interaction between CesT and Map. Similar to other chaperone-effector pairings, binding is apparent at the amino-terminus of Map and is indicated to proceed by a similar mechanism to CesT:Tir interaction. Map secretion from a cesT mutant strain (SE884) is shown to be reduced and, importantly, its translocation from this strain after infection of HEp-2 cells is almost totally abrogated. Although other chaperones are reported to have a bivalent binding specificity, CesT is the first member of its family that chaperones more than one protein for translocation.
authors.affiliation: Centre for Molecular Microbiology and Infection, Department of Biological Sciences, Imperial College, London SW7 2AZ, UK.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Creasey
authors.forename: Robin M
authors.initials: RM
authors.lastname: Delahay
authors.forename: Alexandra A
authors.initials: AA
authors.lastname: Bishop
authors.forename: Robert K
authors.initials: RK
authors.lastname: Shaw
authors.forename: Brendan
authors.initials: B
authors.lastname: Kenny
authors.forename: Stuart
authors.initials: S
authors.lastname: Knutton
authors.forename: Gad
authors.initials: G
authors.lastname: Frankel
mesh_terms: D001692:Biological Transport; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism* / Q000472:pathogenicity; D029968:Escherichia coli Proteins / Q000378:metabolism*; D018832:Molecular Chaperones / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
chemical_list: C122044:CesT protein, E coli; D029968:Escherichia coli Proteins; D018832:Molecular Chaperones; D011956:Receptors, Cell Surface; C109287:Tir protein, E coli