PROTEIN-KINASE-C ACTIVATION BY 12-O-TETRADECANOYLPHORBOL 13-ACETATE MODULATES MESSENGER-RIBONUCLEIC-ACID LEVELS FOR 2 OF THE REGULATORY SUBUNITS OF 3', 5'-CYCLIC ADENOSINE MONOPHOSPHATE-DEPENDENT PROTEIN-KINASES (RII-BETA AND RI-ALPHA) VIA MULTIPLE AND DISTINCT MECHANISMS

Messenger RNAs (mRNA) for two of the regulatory subunits of cAMP-dependent protein kinases (PKA), RII-beta and RI-alpha, are transiently (maximal levels at 6 h) stimulated by 12-O-tetradecanoylphorbol 13-acetate (TPA) in cultured rat Sertoli cells in a time- and concentration-dependent manner. Whereas TPA (10(-7) M) stimulated RII-beta mRNA 11 +/- 2.8 fold (mean +/- SEM), mRNA levels for RI-alpha increased only 2.5 +/- 0.6-fold (mean +/- SEM). No effects of TPA on the other subunits of PKA (RII-alpha, C-alpha) were observed. TPA-dependent accumulation of mRNAs for RII-beta and RI-alpha was observed to the same extent in nucleus and cytoplasm. We have previously shown that mRNA levels for all the PKA subunits are increased by cAMP, particularly that of RII-beta (> 50-fold). TPA modulated the stimulatory effects of cAMP on RII-beta and RI-alpha mRNAs in opposite directions. Whereas treatment with both 8-CPTcAMP and TPA gave an additive effect on RI-alpha mRNA, TPA reduced the cAMP-dependent increase in RII-beta mRNA. Although the mRNA for RII-beta had returned to basal levels after 24 h of incubation with TPA, the presence of TPA still inhibited cAMP-dependent induction of mRNA for RII-beta. In contrast, similar TPA treatment did not influence the subsequent cAMP-dependent stimulation of RI-alpha mRNA. Preincubation with 8-CPTcAMP did not influence TPA-dependent stimulation of mRNAs for either RII-beta or RI-alpha. TPA induction of RII-beta mRNA was completely blocked by cycloheximide (an inhibitor of protein synthesis), whereas that of RI-alpha was not. The inhibitory effect of TPA on cAMP stimulation of RII-beta mRNA was independent of ongoing protein synthesis. These results indicate that TPA induction of mRNAs for RI-alpha and RII-beta involves multiple and distinct mechanisms. The stimulatory effect of TPA on RI-alpha mRNA levels and the inhibitory effect of TPA on cAMP-stimulated RII-beta mRNA expression are probably mediated through stable factors, whereas proteins with rapid turnover or factors induced by TPA are involved in the stimulatory effect of TPA on RII-beta mRNA.