Association of microtubules and intermediate filaments in normal fibroblasts and its disruption upon transformation by a temperature-sensitive mutant of Rous sarcoma virus.

Abstract

By double indirect immunofluorescence using primary rabbit antibodies to tubulin and guinea pig antibodies to vimentin, microtubules and intermediate filaments were simultaneously labeled in serval types of cultured normal fibroblasts. With well-spread interphase cells there was an extensive but not complete correspondence of the labeling patterns for the 2 filamentous structures out to the cell periphery. This correspondence existed at a gross level, where parallel but not coincident arrays of thickly labeled strands of the 2 types of filaments were observed, and at a fine level, where thinly labeled strands of the 2 were superimposed. There may be some type(s) of molecular linkages between microtubules and vimentin intermediate filaments that is under metabolic control. With rat kidney fibroblasts infected with a temperature-sensitive mutant (LA23) of Rous sarcoma virus, cells grown at the nonpermissive temperature (39.degree. C) showed the correspondence of the distributions of the microtubules and intermediate filaments characteristic of the normal phenotype but within 1 h after a shift to the permissive temperature (33.degree. C) there was an extensive retraction of the intermediate filaments around the cell nucleus; the microtubules remained dispersed into the cell periphery. One of the functions carried out by p60src, the protein kinase responsible for transformation by Rous sarcoma virus, may be to modify the component(s) involved in the putative linkages between microtubules and intermediate filaments in the normal cells.