Cytoplasmic localization of the transforming protein of Fujinami sarcoma virus: salt-sensitive association with subcellular components.
Fujinami sarcoma virus (FSV) encodes a transforming protein of 130,000 daltons (P130) which is associated with a tyrosine-specific protein kinase activity. To elucidate mechanisms involved in cell transformation by FSV, we have studied the intracellular location of P130 in rat cells nonproductively infected with FSV. Immunofluorescent staining of several FSV-transformed rat cell lines with a tumor regressor antiserum specific against the fps sequences of P130 showed that the major staining was localized in the cytoplasm. Staining was also seen in cell ruffles and in some cases at areas of cell contact. The cytoplasmic location of P130 staining in cells infected with temperature-sensitive mutants of FSV was unchanged when they were grown at permissive or nonpermissive temperature. Cell fractionation of FSV-transformed cells under various conditions showed that the ionic strength used during cell fractionation had a striking effect on the distribution of P130. At 10 mM NaCl, 70% of P130 sedimented in the large granule fraction, whereas at 500 mM NaCl 70 to 90% of P130 was recovered in the cytosol fraction. Furthermore, a combination of ionic and nonionic detergents that effectively solubilized subcellular membranes was insufficient to solubilize P130 unless the salt concentration was raised. We conclude that the majority of P130 and its associated protein kinase activity are localized in the cytoplasm and that P130 is not an integral membrane protein.
D000818:Animals; D001358:Avian Sarcoma Viruses / Q000032:analysis* / Q000235:genetics; D002478:Cells, Cultured; D003593:Cytoplasm / Q000032:analysis*; D003673:Defective Viruses / Q000032:analysis* / Q000235:genetics; D003902:Detergents; D004622:Embryo, Mammalian; D005347:Fibroblasts; D005455:Fluorescent Antibody Technique; D011494:Protein Kinases / Q000032:analysis*; D011505:Protein-Tyrosine Kinases; D051381:Rats; D012965:Sodium Chloride; D013696:Temperature
D003902:Detergents; D012965:Sodium Chloride; D011494:Protein Kinases; D011505:Protein-Tyrosine Kinases