Tyrosine phosphate hydrolysis of host proteins by an essential Yersinia virulence determinant.
The plasmid-encoded YopH protein is a protein-tyrosine phosphatase (PTPase; EC 3.1.3.48) that is essential for Yersinia virulence. We have investigated the molecular basis for the role of PTPase activity in Yersinia pathogenesis. Allelic recombination was employed to introduce a defined mutation into the yopH plasmid gene. Conversion of the essential Cys-403 to Ala in the catalytic domain of the protein abolished YopH PTPase activity and significantly reduced the virulence of Yersinia pseudotuberculosis in a murine infection model. 32P-labeled phosphotyrosine-containing proteins were immunoprecipitated from extracts of Y. pseudotuberculosis-infected cell monolayers and analyzed by SDS/PAGE to assess the impact of YopH on host protein phosphorylation. Major proteins of 200, 120, and 60 kDa were dephosphorylated in macrophages associated with wild-type Y. pseudotuberculosis. Selective removal of phosphate from the 120- and 60-kDa proteins was shown to be specific to the YopH PTPase activity. Phagocytosis of the bacteria was not required for this dephosphorylation activity, suggesting that YopH is functionally expressed by extracellular bacteria. These observations indicate that the essential function of YopH in Yersinia pathogenesis is host-protein dephosphorylation.
Bliska
J B
JB
Department of Microbiology and Immunology, Stanford University School of Medicine, CA 94305.
Guan
K L
KL
Dixon
J E
JE
Falkow
S
S
D000818:Animals; D002460:Cell Line; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D007700:Kinetics; D008264:Macrophages; D051379:Mice; D016297:Mutagenesis, Site-Directed; D010710:Phosphates / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000235:genetics*; D019000:Phosphotyrosine; D010957:Plasmids; D017027:Protein Tyrosine Phosphatases; D011995:Recombination, Genetic; D015183:Restriction Mapping; D013379:Substrate Specificity; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism; D014774:Virulence; D015011:Yersinia pseudotuberculosis / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*
D010710:Phosphates; D019000:Phosphotyrosine; D014443:Tyrosine; D010749:Phosphoprotein Phosphatases; D017027:Protein Tyrosine Phosphatases