Identification of p130Cas as a substrate of Yersinia YopH (Yop51), a bacterial protein tyrosine phosphatase that translocates into mammalian cells and targets focal adhesions

摘要: A number of pathogenic bacteria utilize type III secretion pathways to translocate virulence proteins into host eukaryotic cells. We identified a target YopH, protein tyrosine phosphatase that is translocated mammalian cells by Yersiniae. catalytically inactive 'substrate-trapping' mutant, YopHC403S, was used as probe determine where YopH substrates localize in Immunofluorescence microscopy demonstrated YopHC403S localized focal adhesions human epithelial infected with Y. pseudotuberculosis. stabilized adhesions, shown its dominant-negative effect on adhesion disassembly mediated YopE, which disrupts actin stress fibers. Conversely, destabilized even the absence loss phosphotyrosine staining. Immunoprecipitation revealed trapped complex hyperphosphorylated 125-135 kDa protein, immunoblotting p130Cas. bound directly p130Cas phosphotyrosine-dependent manner vitro. Translocation plated fibronectin resulted rapid and selective dephosphorylation These results demonstrate targets p130Cas, docking for multiple SH2 domains, direct substrate this enzyme vivo.