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The role of cAMP in the regulation of the amount and synthesis of cytochrome P-450 cholesterol side-chain cleavage (P-450scc) and cytochrome P-450 17 alpha-hydroxylase/C17-20 lyase (P-450(17 alpha) was investigated in mouse Leydig cell cultures. In the absence of cAMP, the amount of immunoreactive P-450(17 alpha) decreased to less than 5% by day 4 and was undetectable between days 7 and 11. In contrast, the amount of immunoreactive P-450scc remained relatively constant throughout the same period. Treatment of Leydig cell cultures for 4 days with 0.05 mM 8-bromo-cAMP initiated on day 7 increased the amount of P-450(17 alpha) with relatively little effect on the amount of P-450scc. The rate of de novo synthesis of each of the P-450 enzymes was studied by determining [35S]methionine incorporation into newly synthesized protein. In the absence of cAMP, de novo synthesis of P-450(17 alpha) ceased while the rate of de novo synthesis of P-450scc increased with time in culture between days 2 and 11. Treatment with cAMP initiated on day 7 of culture caused a time-dependent increase in the rate of de novo synthesis of P-450(17 alpha) on days 9 and 11 equivalent to 40% and 60%, respectively, of that observed in freshly isolated Leydig cells. The rate of de novo synthesis of P-450scc was increased 2-fold relative to untreated cultures on days 9 and 11. De novo synthesis of P-450(17 alpha) ceased when cAMP was removed on day 11 and restored when cAMP was added again on day 13 of culture.(ABSTRACT TRUNCATED AT 250 WORDS)

molecular endocrinology

Noncoordinate Regulation of<i>de Novo</i>Synthesis of Cytochrome P-450 Cholesterol Side-Chain Cleavage and Cytochrome P-450 17α-Hydroxylase/C<sub>17-20</sub>Lyase in Mouse Leydig Cell Cultures: Relation to Steroid Production

Noncoordinate Regulation ofde NovoSynthesis of Cytochrome P-450 Cholesterol Side-Chain Cleavage and Cytochrome P-450 17α-Hydroxylase/C17-20Lyase in Mouse Leydig Cell Cultures: Relation to Steroid Production

Noncoordinate Regulation ofde NovoSynthesis of Cytochrome P-450 Cholesterol Side-Chain Cleavage and Cytochrome P-450 17α-Hydroxylase/C_17-20Lyase in Mouse Leydig Cell Cultures: Relation to Steroid Production