Phosphatidylinositol kinase activity associates with viral p60src protein.
Immunoprecipitates of p60v-src proteins from chicken embryo fibroblasts infected with Rous sarcoma virus were assayed for phosphatidylinositol (PI) kinase activity in the absence of detergents. The product of the PI kinase reaction, phosphatidylinositol monophosphate (PIP), migrated slightly slower than did the authentic phosphatidylinositol-4-monophosphate marker in thin-layer chromatography and was indistinguishable from phosphatidylinositol-3-monophosphate produced by PI kinase type I. Furthermore, the deacylated product comigrated with glycerophosphoinositol-3-phosphate in high-performance liquid chromatography. Both sucrose gradient fractionation and the heat stability of PI kinase activity from cells infected with temperature-sensitive mutants suggest that the PI kinase activity is not intrinsic to p60v-src but is a property of another molecule complexed with p60v-src. All transforming variants of p60src were associated with PI kinase activity, whereas this enzyme activity was hardly detectable in immunoprecipitates from cells infected with nontransforming viruses encoding p60c-src or an enzymatically inactive variant. However, PI kinase activity was found in p60src immunoprecipitates from cells infected with nonmyristylated, nontransforming mutants as well as temperature-sensitive mutants at the nonpermissive temperature, which indicated that simple association of PI kinase activity with p60src is not sufficient for cell transformation.
Fukui
Y
Y
Rockefeller University, New York, New York 10021.
Hanafusa
H
H
D019870:1-Phosphatidylinositol 4-Kinase; D000818:Animals; D001358:Avian Sarcoma Viruses / Q000235:genetics / Q000378:metabolism; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D002642:Chick Embryo; D009154:Mutation; D015688:Oncogene Protein pp60(v-src); D010770:Phosphotransferases / Q000302:isolation & purification / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism; D012191:Retroviridae Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D012191:Retroviridae Proteins; D010770:Phosphotransferases; D011494:Protein Kinases; D019870:1-Phosphatidylinositol 4-Kinase; D015688:Oncogene Protein pp60(v-src)