Cytoskeletal changes induced by two avian sarcoma viruses: UR2 and Rous sarcoma virus

UR2-transformed cells were examined by immunofluorescence and compared to control cells and cells transformed by Rous Sarcoma Virus (RSV). Actin and tubulin which are normally depolymerized in RSV-transformed cells appeared to be unaffected by UR2 transformation. Cell surface fibronectin which is normally lost from RSV-infected cells, appears more abundantly on UR2-transformed cells than on normal cells. Vinculin was shown to be in adhesion plaques in UR2-transformed cells as well as in control fibroblasts but diffuse in the cytoplasm of RSV-transformed cells. Polyacrylamide gel electrophoresis of [35S]methionine-labeled fibronectin and vinculin immunoprecipitated from lysates of normal and transformed cells indicated that cell associated fibronectin synthesized during the labeling period is reduced by 60% in RSV-transformed cells but occurs in normal amounts in UR2-transformed cells. However, immunoprecipitation of radiolabeled fibronectin released in supernatant fluids of normal and transformed cells showed a decreased amount of fibronectin in fluids from UR2-transformed cells, but a considerable increase in the medium from RSV-infected cells as compared to uninfected cultures. These data suggest that more fibronectin binds to the surface of UR2-transformed cells then to normal cells, but is readily released from RSV-transformed cells. Vinculin was reduced by about 50% of normal levels in both RSV- and UR2-transformed cells. Immunofluorescence studies using antibody to virion structural proteins (gag) show that the nuclei of UR2-transformed cells are not fluorescent. This indicates a cytoplasmic location or membrane association for p68ros, the transforming protein of UR2, which contains gag determinants. Overall, these data suggest that changes in the major cytoskeletal proteins of fibroblasts are not essential for the neoplastic properties of cells but are rather a phenotypic expression of transformation, since UR2, which causes tumors in vivo, induces only minor cytoskeletal alterations of cells transformed in vitro.