Autocrine stimulation by the v-sis gene product requires a ligand-receptor interaction at the cell surface

Autocrine expression of a growth factor and its receptor in the same cell raises the possibility of an intracellular receptor-ligand interaction within the cell, in addition to a receptor-ligand interaction at the cell surface. We have constructed a NIH3T3 cell line which contains the v-sis gene under the inducible control of the Drosophila melanogaster hsp70 promoter. Expression of both v-sis RNA and protein is rapidly induced by a short period of heat-shock. We have analyzed the cellular site of interaction between the v-sis protein and the platelet-derived growth factor receptor in these cells. Autophosphorylation of the PDGF receptor and induction of the c-fos gene were found to occur at 45 and 50 min, respectively, after heat-induced synthesis of the v-sis protein. Monensin treatment of the heat-induced cells prevented autophosphorylation of the mature PDGF receptor and also prevented subsequent induction of c-fos. Autophosphorylation of the PDGF receptor and c-fos induction were also prevented by the addition of suramin to the medium. These results demonstrate that autocrine stimulation, as monitored by c-fos induction and by PDGF receptor autophosphorylation, requires an interaction between the v-sis protein and the PDGF receptor that occurs at the cell surface, rather than an intracellular location.