Abstract
The regulation of the genes encoding cholesterol side-chain cleavage enzyme (P450scc) and ferredoxin, two components in the first step of steroid synthetic pathways, was studied by RNA analyses of endogenous and transfected genes. cAMP rather than calcium was the major secondary messenger that stimulated expression of both P450scc and ferredoxin genes in human placental JEG-3 cells. The effect of cAMP on P450scc expression was abolished by cycloheximide in JEG-3 cells, but it was superinduced in mouse adrenal Y1 cells. For ferredoxin expression, both reagents have synergistic effect in Y1 and JEG-3 cells. To test the mechanism of regulation, DNA segments containing regulatory elements of the P450scc and ferredoxin genes were connected to reporter genes and analyzed in cotransfection experiments. The results showed that the proximal cAMP-responsive sequences of both P450scc and ferredoxin genes were stimulated by cAMP early in both Y1 and JEG-3 cells, requiring no new protein synthesis. This indicates a common mechanism for the regulated expression of both genes. P450scc possessed an additional upstream cAMP-responsive sequence that also responded to cAMP induction in a different manner from the proximal element. The presence of additional upstream regulatory elements makes it possible for the P450scc gene to be further regulated.
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