title: Sequential disassembly of the cytoskeleton in BHK21 cells infected with vesicular stomatitis virus.
abstract: The cytopathic effects of vesicular stomatitis virus (VSV) that result in the rounding of BHK21 cells have been studied. The results indicate that they are mediated by a sequential alteration in the distribution of the components of the cytoskeleton, an effect that requires the expression of the viral L protein. The constituents of the cytoskeleton of BHK21 cells were analyzed by fluorescence microscopy. Actin filaments were the first component to become disorganized, so that disassembly of stress fibers were detected 1 hr after infection. The distribution of microtubules and intermediate filaments was unchanged at 2 hr after infection; however, both these cytoskeletal elements exhibited an altered distribution at 3-4 hr after infection. Actinomycin D and cycloheximide did not cause the same effects as infection with VSV, suggesting that inhibition of host-cell gene expression was not responsible. However, viral gene expression was required, since cells infected with uv-irradiated VSV showed the same distribution of cytoskeletal constituents as mock-infected controls. Cells infected at 39.5 degrees (the nonpermissive temperature) with mutants of VSV temperature sensitive in the viral NS (ts G22), N(ts G41), M(ts 0 23), and G(ts 0 45) proteins showed the same changes in the cytoskeleton as those detected with wild-type virus. In contrast, cells infected with ts G11 (L-) showed the characteristic effect of VSV on the cytoskeleton when incubated at 34 degrees (the permissive temperature), but not when incubated at 39.5 degrees. The T-1026 R1 mutant of VSV, which has a much less dramatic effect on cell morphology than wild-type virus, also caused a less marked disruption of the cytoskeleton.
authors.affiliation: Department of Neurobiology, Anatomy, and Cell Science, University of Pittsburgh School of Medicine, Pennsylvania 15261.
authors.forename: K O
authors.initials: KO
authors.lastname: Simon
authors.forename: P A
authors.initials: PA
authors.lastname: Whitaker-Dowling
authors.forename: J S
authors.initials: JS
authors.lastname: Youngner
authors.forename: C C
authors.initials: CC
authors.lastname: Widnell
mesh_terms: D000818:Animals; D002460:Cell Line; D002472:Cell Transformation, Viral; D006224:Cricetinae; D003513:Cycloheximide / Q000494:pharmacology; D003598:Cytoskeletal Proteins / Q000032:analysis; D003599:Cytoskeleton / Q000187:drug effects / Q000648:ultrastructure*; D003609:Dactinomycin / Q000494:pharmacology; D015870:Gene Expression; D007382:Intermediate Filaments / Q000648:ultrastructure; D007668:Kidney; D009154:Mutation; D014721:Vesicular stomatitis Indiana virus / Q000187:drug effects / Q000235:genetics*
chemical_list: D003598:Cytoskeletal Proteins; D003609:Dactinomycin; D003513:Cycloheximide