The amino-terminal 14 amino acids of v-src can functionally replace the extracellular and transmembrane domains of v-erbB.
The retroviral oncogene v-erbB encodes a truncated form of the receptor for epidermal growth factor, an integral membrane protein-tyrosine kinase. By contrast, the oncogene v-src encodes a protein-tyrosine kinase that is a peripheral membrane protein. The morphologies and spectra of cells transformed by these two oncogenes differ. In an effort to identify the functional determinant(s) of these differences, we constructed and tested first deletion mutants of v-erbB and then chimeras between v-src and v-erbB. As reported previously, the absence of any membrane anchorage eliminated transformation by v-erbB. Anchorage of the cytoplasmic kinase domain of v-erbB to membranes with amino-terminal portions of the v-src protein permitted transformation. The phenotype and spectrum of transformation were those expected for v-erbB rather than for v-src. The transforming chimeras lost their biological activity if the signal for myristylation at the amino terminus of v-src was compromised by mutation. Biochemical fractionations revealed a correlation between transforming activity and the association of chimeric gene products with the membrane fraction of the cell. For reasons not yet apparent, the combined presence of membrane anchorage domains of v-src, and the transmembrane domain of v-erbB in the same chimera typically (but not inevitably) impeded transformation. Our results suggest that the specificity of transformation by v-erbB resides in the selection of substrates by the cytoplasmic domain of the gene product. The protein retains access to those substrates even when anchored to the membrane in the manner of a peripheral rather than a transmembrane protein.
Department of Microbiology and Immunology, University of California, San Francisco 94143.
M
M
McMahon
R C
RC
Schatzman
J M
JM
Bishop
D000483:Alleles; D000818:Animals; D015153:Blotting, Western; D002458:Cell Fractionation; D002460:Cell Line; D002471:Cell Transformation, Neoplastic / Q000235:genetics; D002478:Cells, Cultured; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051379:Mice; D015688:Oncogene Protein pp60(v-src) / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018739:Oncogene Proteins v-erbB; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015682:Retroviridae Proteins, Oncogenic / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D008565:Membrane Proteins; D018739:Oncogene Proteins v-erbB; D011993:Recombinant Fusion Proteins; D015682:Retroviridae Proteins, Oncogenic; D015688:Oncogene Protein pp60(v-src)