Monoclonal antibodies to the p21 products of the transforming gene of Harvey murine sarcoma virus and of the cellular ras gene family.
We have isolated eight rat lymphocyte-myeloma hybrid cell lines producing monoclonal antibodies that react with the 21,000-dalton transforming protein (p21) encoded by the v-ras gene of Harvey murine sarcoma virus (Ha-MuSV). These antibodies specifically immunoprecipitate both phosphorylated and non-phosphorylated forms of p21 from lysates of cells transformed by Ha-MuSV. All eight react with the products of closely related ras genes expressed in cells transformed by two additional sarcoma viruses (rat sarcoma virus and BALB sarcoma virus) or by a cellular Harvey-ras gene placed under the control of a viral promoter. Three of the antibodies also react strongly with the p21 encoded by the v-ras gene of Kirsten MuSV. These same three antibodies immunoprecipitate the predominant p21 species synthesized normally in a variety of rodent cell lines, including the p21 produced at high levels in 416B murine hemopoietic cells. This suggests that an endogenous gene closely related to Kirsten-ras is expressed in these cells. The monoclonal antibodies have been used to confirm two properties associated with p21; localization at the inner surface of the membrane of Ha-MuSV-transformed cells, assayed by immunofluorescence microscopy, and binding of guanine nucleotides.
D000818:Animals; D000911:Antibodies, Monoclonal; D002460:Cell Line; D002462:Cell Membrane / Q000032:analysis*; D002471:Cell Transformation, Neoplastic; D002472:Cell Transformation, Viral; D005814:Genes, Viral; D006153:Guanosine Diphosphate / Q000378:metabolism; D051379:Mice; D009858:Oncogenic Viruses / Q000235:genetics; D051381:Rats; D009053:Sarcoma Viruses, Murine / Q000032:analysis* / Q000235:genetics; D014764:Viral Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism
D000911:Antibodies, Monoclonal; D014764:Viral Proteins; D006153:Guanosine Diphosphate