Isolation of temperature-sensitive Abelson virus mutants by site-directed mutagenesis.
Mutants of Abelson virus encoding temperature-sensitive protein-tyrosine kinase (EC 2.7.1.112) were created by site-directed mutagenesis using sequence information from temperature-sensitive mutants of the related v-src oncogene. Expression of these two independent mutations in Escherichia coli resulted in reduced phosphorylation of the mutant proteins at high temperature. Viruses containing one of the mutations induced conditional transformation of both NIH 3T3 and lymphoid cells when expressed in the context of a truncated transforming protein. These results underscore the functional homology between protein-tyrosine kinases and suggest that transfer of mutations within a related gene family may provide a rapid method to create mutants.
Engelman
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A
Department of Molecular Biology, Tufts University School of Medicine, Boston, MA 02111.
Rosenberg
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N
D000011:Abelson murine leukemia virus / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002472:Cell Transformation, Viral; D005347:Fibroblasts; D009052:Leukemia Virus, Murine / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000235:genetics* / Q000378:metabolism; D012191:Retroviridae Proteins / Q000235:genetics* / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D013696:Temperature; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism
D012191:Retroviridae Proteins; D014764:Viral Proteins; D011505:Protein-Tyrosine Kinases