Identification of CesT, a chaperone for the type III secretion of Tir in enteropathogenic Escherichia coli.
The locus of enterocyte effacement of enteropathogenic Escherichia coli encodes a type III secretion system, an outer membrane protein adhesin (intimin, the product of eae ) and Tir, a translocated protein that becomes a host cell receptor for intimin. Many type III secreted proteins require chaperones, which function to stabilize proteins, prevent inappropriate protein-protein interactions and aid in secretion. An open reading frame located between tir and eae, previously named orfU, was predicted to encode a protein with partial similarity to the Yersinia SycH chaperone. We examined the potential of the orfU gene product to serve as a chaperone for Tir. The orfU gene encoded a 15 kDa cytoplasmic protein that specifically interacted with Tir as demonstrated by the yeast two-hybrid assay, column binding and coimmunoprecipitation experiments. An orfU mutant was defective in attaching-effacing lesion formation and Tir secretion, but was unaffected in expression of other virulence factors. OrfU appeared to stabilize Tir levels in the cytoplasm, but was not absolutely necessary for secretion of Tir. Based upon the physical similarities, phenotypic characteristics and the demonstrated interaction with Tir, orfU is redesignated as cesT for the chaperone for E. coli secretion of T ir.
Elliott
S J
SJ
Center for Vaccine Development and Department of Microbiology and Immunology, University of Maryland School of Medicine, 685 W Baltimore St, Baltimore, MD 21201, USA.
Hutcheson
S W
SW
Dubois
M S
MS
Mellies
J L
JL
Wainwright
L A
LA
Batchelor
M
M
Frankel
G
G
Knutton
S
S
Kaper
J B
JB
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology*; D001483:Base Sequence; D003001:Cloning, Molecular; D017124:Conserved Sequence; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology*; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial; D018832:Molecular Chaperones / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000235:genetics / Q000502:physiology*; D017386:Sequence Homology, Amino Acid; D012760:Shigella / Q000235:genetics / Q000502:physiology; D014170:Transformation, Genetic; D014774:Virulence / Q000235:genetics / Q000502:physiology
D001426:Bacterial Proteins; C122044:CesT protein, E coli; D017931:DNA Primers; D029968:Escherichia coli Proteins; D018832:Molecular Chaperones; D011956:Receptors, Cell Surface; C109287:Tir protein, E coli