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

被引:105
作者
Gauthier, A
Finlay, BB [1 ]
机构
[1] Univ British Columbia, Biotechnol Lab, Vancouver, BC V6T 1Z3, Canada
[2] Univ British Columbia, Dept Biochem, Vancouver, BC V6T 1Z3, Canada
[3] Univ British Columbia, Dept Microbiol, Vancouver, BC V6T 1Z3, Canada
关键词
D O I
10.1128/JB.185.23.6747-6755.2003
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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 FIR 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.
引用
收藏
页码:6747 / 6755
页数:9
相关论文
共 61 条
[21]   Secretin of the enteropathogenic Escherichia coli type III secretion system requires components of the type III apparatus for assembly and localization [J].
Gauthier, A ;
Puente, JL ;
Finlay, BB .
INFECTION AND IMMUNITY, 2003, 71 (06) :3310-3319
[22]   Enteropathogenic E-coli translocated intimin receptor, Tir, interacts directly with α-actinin [J].
Goosney, DL ;
DeVinney, R ;
Pfuetzner, RA ;
Frey, EA ;
Strynadka, NC ;
Finlay, BB .
CURRENT BIOLOGY, 2000, 10 (12) :735-738
[23]   Role of the cytoplasmic C terminus of the FliF motor protein in flagellar assembly and rotation [J].
Grünenfelder, B ;
Gehrig, S ;
Jenal, U .
JOURNAL OF BACTERIOLOGY, 2003, 185 (05) :1624-1633
[24]   THE BINDING CASCADE OF SECB TO SECA TO SECY/E MEDIATES PREPROTEIN TARGETING TO THE ESCHERICHIA-COLI PLASMA-MEMBRANE [J].
HARTL, FU ;
LECKER, S ;
SCHIEBEL, E ;
HENDRICK, JP ;
WICKNER, W .
CELL, 1990, 63 (02) :269-279
[25]   Interactions between type III secretion apparatus components from Yersinia pestis detected using the yeast two-hybrid system [J].
Jackson, MW ;
Plano, GV .
FEMS MICROBIOLOGY LETTERS, 2000, 186 (01) :85-90
[26]   Enteropathogenic Escherichia coli (EPEC) -: a crafty subversive little bug [J].
Kenny, B .
MICROBIOLOGY-SGM, 2002, 148 :1967-1978
[27]   Enteropathogenic Escherichia coli (EPEC) Tir receptor molecule does not undergo full modification when introduced into host cells by EPEC-independent mechanisms [J].
Kenny, B ;
Warawa, J .
INFECTION AND IMMUNITY, 2001, 69 (03) :1444-1453
[28]   Enteropathogenic E-coli (EPEC) transfers its receptor for intimate adherence into mammalian cells [J].
Kenny, B ;
DeVinney, R ;
Stein, M ;
Reinscheid, DJ ;
Frey, EA ;
Finlay, BB .
CELL, 1997, 91 (04) :511-520
[29]   Deletion analysis of the flagellar switch protein FliG of Salmonella [J].
Kihara, M ;
Miller, GU ;
Macnab, RM .
JOURNAL OF BACTERIOLOGY, 2000, 182 (11) :3022-3028
[30]   Pas, a novel protein required for protein secretion and attaching and effacing activities of enterohemorrhagic Escherichia coli [J].
Kresse, AU ;
Schulze, K ;
Deibel, C ;
Ebel, F ;
Rohde, M ;
Chakraborty, T ;
Guzmán, CA .
JOURNAL OF BACTERIOLOGY, 1998, 180 (17) :4370-4379