Post-Translational Regulation of Steroidogenic Acute Regulatory Protein by cAMP-Dependent Protein Kinase A

Abstract

Adrenal steroid production is stimulated by adrenocorticotropin hormone activation of the cAMP-dependent protein kinase A (PKA) signaling pathway and subsequent induction of Steroidogenic Acute Regulatory (StAR) protein expression. Herein we have compared StAR mRNA and protein levels in 8-Br-cAMP-treated mouse adrenocortical Y1 and the derived PKA mutant Kin-8 cell lines to evaluate the PKA requirement in StAR expression. StAR mRNA was induced by 8-Br-cAMP-treatment of both Y1 and Kin-8 cells with maximal expression levels in Kin-8 cells approximately 50% of that observed in Y1 cells. StAR protein levels, as detected by Western analysis, were concomitantly increased in Y1 cells but were not detected in the Kin-8 cells. StAR mRNA colocalized with the active polysome fractions in both 8-Br-cAMP-treated Y1 and Kin-8 cells, indicating translation was not blocked in Kin-8 cells. Consistent with this data, a 2-fold increase in incorporation of [35S]methionine into StAR was also observed after 8-Br-cAMP treatment of both cell lines. Since StAR protein levels were not sufficient to detect by Western analysis, these data indicate that PKA functions at the post-translational level to regulate StAR expression and we propose that phosphorylation of StAR by PKA contributes to protein stability.