Recombinant Yersinia YopT Leads to Uncoupling of RhoA-Effector Interaction

AUTOR(ES)

Sorg, Isabel

FONTE

American Society for Microbiology

RESUMO

Yersinia enterocolitica, Yersinia pseudotuberculosis, and Yersinia pestis deliver different Yop (Yersinia outer proteins) effector proteins into mammalian cells by a type III secretion mechanism. Recently, it was shown that Yersinia producing YopT leads to disruption of the actin cytoskeleton of HeLa cells (M. Iriarte and G. R. Cornelis, Mol. Microbiol. 29:915–929, 1998). To analyze the molecular mechanism of YopT, we cloned and expressed YopT as a glutathione S-transferase fusion protein. Recombinant YopT caused rounding up of embryonic bovine lung cells and redistribution of the actin cytoskeleton rapidly after microinjection. The Escherichia coli cytotoxic necrotizing factor (CNF1), which constitutively activates Rho proteins, was not able to inhibit or revert YopT-induced cell rounding. YopT caused release of RhoA from embryonic bovine lung membranes and released recombinant isoprenylated RhoA from artificial PE or PE/PIP2 vesicles. Incubation of lysate or cytosol with YopT caused inhibition of the RhoA-rhotekin interaction but led to increased RhoA-RhoGDI interaction. It is suggested that inhibition of the interaction between RhoA and effectors is the underlying mechanism of the YopT action on the cytoskeleton.

Documentos Relacionados

• Yersinia pseudotuberculosis Virulence Determinants Invasin, YopE, and YopT Modulate RhoG Activity and Localization▿
• Biochemical characterization of the Yersinia YopT protease: Cleavage site and recognition elements in Rho GTPases
• The C Terminus of YopT Is Crucial for Activity and the N Terminus Is Crucial for Substrate Binding
• The Yersinia YopE and YopH type III effector proteins enhance bacterial proliferation following contact with eukaryotic cells
• LcrG is Required for Efficient Translocation of Yersinia Yop Effector Proteins into Eukaryotic Cells