Interaction between tubulin and the viral matrix protein of vesicular stomatitis virus: possible implications in the viral cytopathic effect.
The matrix (M) protein of vesicular stomatitis virus has been shown to induce the rounding of cells. Experiments were performed in order to define the mechanism by which M protein could cause this cytopathic effect (CPE). Immunofluorescence experiments performed on infected cells indicate that cellular rounding coincides with the disruption of the microtubular network. Immunoprecipitation of M protein or tubulin in infected cell extract demonstrates an association of these two proteins in vivo. We show that M protein is capable of interacting in vitro with tubulin in both its polymerized and nonassembled forms. Studies using proteolytically cleaved proteins indicate that this interaction occurs via the highly basic N-terminal domain of M protein and the highly acidic C-terminal region of tubulin. Furthermore, a thermosensitive mutant (tsG33) containing a mutation in the matrix protein gene which is unable to induce CPE at nonpermissive temperature interacts with tubulin with a lower affinity. These results demonstrate that M protein interacts with tubulin in vivo and in vitro and strongly suggest that CPE is caused by this interaction.
Melki
R
R
Laboratoire d'enzymologie, CNRS, Gif Sur Yvette, France.
Gaudin
Y
Y
Blondel
D
D
D000818:Animals; D002460:Cell Line; D003101:Collodion; D006224:Cricetinae; D003588:Cytopathogenic Effect, Viral; D005455:Fluorescent Antibody Technique; D008567:Membranes, Artificial; D008870:Microtubules / Q000378:metabolism / Q000648:ultrastructure; D011233:Precipitin Tests; D013381:Subtilisins / Q000494:pharmacology; D013696:Temperature; D014404:Tubulin / Q000378:metabolism*; D014721:Vesicular stomatitis Indiana virus / Q000378:metabolism* / Q000472:pathogenicity; D014763:Viral Matrix Proteins / Q000378:metabolism*
D008567:Membranes, Artificial; D014404:Tubulin; D014763:Viral Matrix Proteins; D003101:Collodion; D013381:Subtilisins