Mutagenic analysis of the v-crk oncogene: requirement for SH2 and SH3 domains and correlation between increased cellular phosphotyrosine and transformation.
We have constructed a series of mutants with deletion, linker insertion, and point mutations in the v-crk oncogene of avian sarcoma virus CT10. The v-crk gene contains no apparent catalytic domain, but does contain two blocks of homology to putative regulatory domains, termed SH2 and SH3, found in a variety of proteins implicated in signal transduction. Infection with CT10 causes a dramatic increase in the level of tyrosine phosphorylation of several cellular proteins. We found that mutation of either the SH2 or SH3 domain of v-crk reduced or eliminated transforming activity, whereas mutation of regions outside the conserved domains had no effect. Deletion of amino-terminal gag sequences caused a partial loss of transforming activity and a change in subcellular distribution of the crk protein. In all cases, there was an absolute correlation between increased cellular phosphotyrosine and transformation.
Mayer
B J
BJ
Laboratory of Molecular Oncology, Rockefeller University, New York, New York 10021.
Hanafusa
H
H
D000818:Animals; D001358:Avian Sarcoma Viruses / Q000235:genetics*; D001483:Base Sequence; D002448:Cell Adhesion; D002455:Cell Division; D002471:Cell Transformation, Neoplastic; D002642:Chick Embryo; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D015345:Oligonucleotide Probes; D050736:Oncogene Protein v-crk; D009857:Oncogenes; D010750:Phosphoproteins / Q000302:isolation & purification; D019000:Phosphotyrosine; D010957:Plasmids; D015183:Restriction Mapping; D015682:Retroviridae Proteins, Oncogenic / Q000235:genetics*; D014443:Tyrosine / Q000031:analogs & derivatives / Q000032:analysis
D015345:Oligonucleotide Probes; D050736:Oncogene Protein v-crk; D010750:Phosphoproteins; D015682:Retroviridae Proteins, Oncogenic; D019000:Phosphotyrosine; D014443:Tyrosine