BRYOSTATIN 1 PROTECTS PROTEIN KINASE-C-DELTA FROM DOWN-REGULATION IN MOUSE KERATINOCYTES IN PARALLEL WITH ITS INHIBITION OF PHORBOL ESTER-INDUCED DIFFERENTIATION

Bryostatin 1 and phorbol-12-myristate-13-acetate (PMA) are both potent activators of protein kinase C (PKC), although in primary mouse kerat inocytes bryostatin 1 does not induce differentiation and blocks PMA-i nduced differentiation. We report here that in primary mouse keratinoc ytes PMA caused translocation of PKC-epsilon to the Triton X-100-solub le fraction with an approximately 2-order of magnitude higher potency, compared with translocation of PKC-alpha and PKC-delta. The kinetics of translocation were fastest for PKC-epsilon, slower for PKC-alpha, a nd slowest for PKC-delta. At 5-20 min bryostatin 1 showed potency simi lar to that of PMA for translocating PKC-alpha, higher potency for tra nslocating PKC-delta, and lower potency for translocating PKC-epsilon. At a later time (6 hr), bryostatin 1 was 1-2 orders magnitude more po tent than PMA for causing loss of PKC-alpha, -delta, and -epsilon from the soluble fraction. Bryostatin 1 was 40-fold more potent than PMA f or down-regulating PKC-alpha and showed a biphasic dose-response curve for down-regulating PKC-delta. Bryostatin 1 at 0.1-1 nM down-regulate d PKC-delta to a similar extent as did PMA. Bryostatin 1 at 100 nM to 1 mu M, on the other hand, failed to induce down-regulation, and these high (100 nM to 1 mu M) doses of bryostatin 1 showed noncompetitive i nhibition of PKC-delta down-regulation by 1 mu M PMA after coapplicati on. This protected portion of PKC-delta retained kinase activity. The dose-response curve for bryostatin 1 protection of PKC-delta from down -regulation by PMA correlated with bryostatin 1 inhibition of the effe cts of PMA on cornified envelope formation (a marker of differentiatio n) and epidermal growth factor binding. Although PKC-epsilon was readi ly translocated by both PMA and bryostatin 1, the PKC-epsilon original ly associated with the particulate fraction showed no down-regulation by either of these agents. We hypothesize that differential regulation of PKC isozymes by PMA and bryostatin 1 may contribute to the differe nt patterns of biological responses that they induce.