THE ROLES OF CAMP AND CAMP-DEPENDENT PROTEIN-KINASE IN THE EXPRESSION OF CHOLESTEROL SIDE-CHAIN CLEAVAGE AND STEROID 11-BETA-HYDROXYLASE GENES IN MOUSE ADRENOCORTICAL TUMOR-CELLS
- 5 August 1989
- journal article
- research article
- Vol. 264 (22) , 12867-12871
Abstract
The expression of the genes encoding cholesterol side chain cleavage enzyme (SCC) and steroid 11.beta.-hydroxylase (11.beta.-OHase) was examined in Y1 mouse adrenocortical tumor cells and in derivative cell lines defective in cAMP-dependent protein kinase activity. Y1 cells expressed both genes constitutively, and treatment with 8-bromo-cAMP (8-Br-cAMP) increased expression 5-10-fold. In three independent protein kinase mutants, expression of SCC and 11.beta.-OHase was impaired to degrees dependent upon the severity of defect in cAMP-dependent protein kinase activity. In Kin-2, the least impaired mutant clone, basal expression of SCC was the same as in Y1 cells. Treatment of Kin-2 with 8-Br-cAMP increased SCC RNA to the levels seen in stimulated Y1 cells. In contrast, clone Kin-8, the most severe mutant, expressed markedly diminished basal and 8-Br-cAMP-stimulated levels of SCC mRNA. Kin-7 had basal and 8-Br-cAMP-stimulated levels of SCC mRNA which were intermediate to Kin-2 and Kin-8. None of the Kin mutants constitutively expressed detectable levels of 11.beta.-OHase transcripts, and only Kin-2 responded to treatment with 8-Br-cAMP with increased expression of 11.beta.-OHase; however, the time course of induction in Kin-2 was significantly delayed. The disparate patterns of expression of SCC and 11.beta.-OHase in the Kin mutants suggest that these genes differ in their absolute requirement for cAMP-dependent protein kinase activity. Experiments also were performed in which Kin-7 and Kin-8 mutants were restored to cAMP-responsive states by transfection with genes encodoing normal subunits of cAMP-dependent protein kinase. These phenotypic revertants recovered 8-Br-cAMP-inducible expression of SCC and 11.beta.-OHase. These results strongly support the hypothesis that impaired expression of steroidogenic enzymes in the Kin mutants results directly from defects in cAMP-dependent protein kinase activity.This publication has 18 references indexed in Scilit:
- Cholesterol Side-Chain Cleavage P450 Messenger Ribonucleic Acid: Evidence for Hormonal Regulation in Rat Ovarian Follicles and Constitutive Expression in Corpora Lutea*Endocrinology, 1987
- The roles of cAMP and cAMP-dependent protein kinase in the regulation of adrenocortical functions: analysis using DNA-mediated gene transferBiochemistry and Cell Biology, 1986
- Human growth hormone as a reporter gene in regulation studies employing transient gene expression.Molecular and Cellular Biology, 1986
- Identification of a cyclic-AMP-responsive element within the rat somatostatin gene.Proceedings of the National Academy of Sciences, 1986
- Recovery of hormonal regulation in protein kinase defective adrenal cells through DNA‐mediated gene transferJournal of Cellular Physiology, 1986
- ALTERATION OF THE REGULATORY SUBUNIT OF TYPE-1 CAMP-DEPENDENT PROTEIN-KINASE IN MUTANT Y1 ADRENAL-CELLS RESISTANT TO 8-BROMOADENOSINE 3'-5'-MONOPHOSPHATE1982
- Regulation of ornithine decarboxylase activity by corticotropin in adrenocortical tumor cell clones: roles of cyclic AMP and cyclic AMP-dependent protein kinase.Proceedings of the National Academy of Sciences, 1980
- Isolation of biologically active ribonucleic acid from sources enriched in ribonucleaseBiochemistry, 1979
- Mutations in cyclic AMP-dependent protein kinase and corticotropin (ACTH)-sensitive adenylate cyclase affect adrenal steroidogenesis.Proceedings of the National Academy of Sciences, 1979
- CLONAL ANALYSIS OF DIFFERENTIATED FUNCTION IN ANIMAL CELL CULTURES .I. POSSIBLE CORRELATED MAINTENANCE OF DIFFERENTIATED FUNCTION AND DIPLOID KARYOTYPE1966