A short sequence in the p60src N terminus is required for p60src myristylation and membrane association and for cell transformation.
We have constructed mutants by using linker insertion followed by deletion in the region of cloned Rous sarcoma virus DNA coding for the N-terminal 9 kilodaltons of the src protein. Previous work implicated this region in the membrane association of the protein. The mutations had little effect on src tyrosine kinase activity. Substitution of a tri- or tetrapeptide for amino acids 15 to 27, 15 to 49, or 15 to 81 had little effect on the in vitro transforming capacity of the virus. Like wild-type p60src, the src proteins of these mutants associated with plasma membranes and were labeled with [3H]myristic acid. In contrast, a mutant whose src protein had the dipeptide Asp-Leu substituted for amino acids 2 to 81 and a mutant with the tripeptide Asp-Leu-Gly substituted for amino acids 2 to 15 were transformation defective, and the mutant proteins did not associate with membranes and were not labeled with [3H]myristic acid. These results suggest that amino acids 2 to 15 serve as an attachment site for myristic acid and as a membrane anchor. Since deletions including this region prevent transformation, and since tyrosine kinase activity is not diminished by the deletions, these results imply that target recognition is impaired by mutations altering the very N terminus, perhaps through their effect on membrane association.
D000595:Amino Acid Sequence; D000818:Animals; D001358:Avian Sarcoma Viruses / Q000235:genetics*; D001483:Base Sequence; D002471:Cell Transformation, Neoplastic; D002642:Chick Embryo; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D005347:Fibroblasts; D005796:Genes; D005814:Genes, Viral; D009154:Mutation; D015688:Oncogene Protein pp60(v-src); D010957:Plasmids; D011494:Protein Kinases / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics* / Q000302:isolation & purification
D014764:Viral Proteins; D011494:Protein Kinases; D015688:Oncogene Protein pp60(v-src); D004262:DNA Restriction Enzymes