A mutation in the catalytic domain of pp60v-src is responsible for the host- and temperature-dependent phenotype of the Rous sarcoma virus mutant tsLA33-1.
We have analyzed a host- and temperature-dependent mutant of Rous sarcoma virus in order to learn more about the nature of mutations which lead to a host range phenotype. We have cloned and sequenced the v-src genes from this mutant, tsLA33-1, and from its presumed parent, tsLA33. Both the tsLA33 and the tsLA33-1 pp60v-src proteins contain multiple mutations. The tsLA33 v-src gene product has amino acid alterations at four positions. In the tsLA33-1 v-src gene product, two of these four mutations have reverted to wild type. We have constructed chimeras between the two mutant v-src gene products and between each mutant and the Prague A v-src gene product. To assess the contribution of each amino acid change to the transformation phenotypes of tsLA33 and tsLA33-1, we expressed the hybrid proteins in both chicken embryo fibroblasts and Rat-3 fibroblasts. Additionally, we have measured the protein tyrosine kinase activity of chimeras constructed between the tsLA33 and tsLA33-1 pp60v-src proteins. Our results indicate that mutations in the catalytic domain of each protein are the principal determinants of the transforming ability and protein tyrosine kinase activity of the tsLA33 and tsLA33-1 pp60v-src proteins.
Foster
R
R
Department of Molecular and Cell Biology, University of California, Berkeley 94720.
Martin
G S
GS
D000818:Animals; D001358:Avian Sarcoma Viruses / Q000235:genetics*; D001665:Binding Sites; D002472:Cell Transformation, Viral / Q000235:genetics*; D002478:Cells, Cultured; D002642:Chick Embryo; D003001:Cloning, Molecular; D015151:Immunoblotting; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D015688:Oncogene Protein pp60(v-src) / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013696:Temperature
D011993:Recombinant Fusion Proteins; D015688:Oncogene Protein pp60(v-src)