Bombesin, platelet-derived growth factor, and diacylglycerol induce selective membrane association and down-regulation of protein kinase C isotypes in Swiss 3T3 cells.

Swiss 3T3 cells contain protein kinase C (PKC) isotypes alpha, delta, epsilon and zeta (Olivier, A. R., and Parker, P. J. (1992) J. Cell. Physiol. 152, 240-244). Acute stimulation of quiescent cells with the neuropeptide bombesin decreases the mobility of PKC-delta and PKC-epsilon on SDS-polyacrylamide gels. These slower migrating forms of PKC-delta and PKC-epsilon rapidly (within 1 s) and selectively are found associated with the Triton X-100-soluble membrane fraction. No change in the mobility or distribution of PKC-alpha or PKC-zeta is detected. Long-term treatment of cells with bombesin induces selective membrane association and down-regulation of PKC-delta and PKC-epsilon (decreasing 70 and 65%, respectively). No change in the long-term distribution of PKC-alpha and PKC-zeta was detected. Bombesin did, however, increase PKC-alpha protein levels by 60% compared to control cells. PKC-zeta levels remained unchanged. Both the shift in mobility and down-regulation of PKC-delta and PKC-epsilon were only induced by mitogenic doses of bombesin. The potent mitogen platelet-derived growth factor induced similar effects on the PKC isotypes delta and epsilon. PKC-alpha and PKC-zeta levels were unaffected. Repeated doses of the synthetic diglyceride 1-oleoyl-2-acetyl-sn-glycerol induced PKC-delta and PKC-epsilon down-regulation and stimulated the cells to divide. Again PKC-alpha and PKC-zeta levels were unaffected. These results show a correlation between the membrane association and down-regulation of PKC-delta and PKC-epsilon and the entry of cells into S phase.