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Raikhinstein, M., Department of Hormone Research, Weizmann Institute of Science, Rehovot, 76100, Israel
Hanukoglu, I., Department of Hormone Research, Weizmann Institute of Science, Rehovot, 76100, Israel
To search for genes that are induced by ACTH in adrenocortical cells, we screened adrenal cortex cDNA libraries by a differential hybridization method using cDNA probes representing mRNAs from cells with or without ACTH stimulation. Forty clones were identified as ACTH induced (yielding a frequency of about 1/2500 plaques screened), and two clones as aCTH repressed. The cDNAs isolated and sequenced include nuclear genes for microsomal steroidogenic enzymes and novel proteins of yet unidentified functions, and mitochondrial genes encoding subunits of oxidative phosphorylation enzymes. Northern blot analysis of RNA from cells stimulated with ACTH confirmed the induction of these genes by ACTH, yet revealed important differences in the relative responses of the respective mRNAs. The time courses showed the major increase in the initial 6 h; and a decline after 24-36 h. The enhancement of the levels of the mRNAs could be ascribed to transcriptional activation. Since the mitochondrial genome is transcribed as a single polycistronic unit, to account for the > 20-fold differences in the levels of the mitochondrial mRNAs it is necessary to invoke differential stabilities of these mRNAs. The synchronous increase in the expression of both the steroidogenic enzymes and the mitochondrial oxidative phosphorylation system subunits, provides evidence for coregulation of steroidogenic and energy producing capacities of adrenal cells to meet the metabolic needs of steroid hormone production. Suppression of β-actin gene expression may be related to changes in actin polymerization during ACTH-dependent cytoskeletal reorganization. © 1994.
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Cloning of ACTH-regulated genes in the adrenal cortex
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Raikhinstein, M., Department of Hormone Research, Weizmann Institute of Science, Rehovot, 76100, Israel
Hanukoglu, I., Department of Hormone Research, Weizmann Institute of Science, Rehovot, 76100, Israel
Cloning of ACTH-regulated genes in the adrenal cortex
To search for genes that are induced by ACTH in adrenocortical cells, we screened adrenal cortex cDNA libraries by a differential hybridization method using cDNA probes representing mRNAs from cells with or without ACTH stimulation. Forty clones were identified as ACTH induced (yielding a frequency of about 1/2500 plaques screened), and two clones as aCTH repressed. The cDNAs isolated and sequenced include nuclear genes for microsomal steroidogenic enzymes and novel proteins of yet unidentified functions, and mitochondrial genes encoding subunits of oxidative phosphorylation enzymes. Northern blot analysis of RNA from cells stimulated with ACTH confirmed the induction of these genes by ACTH, yet revealed important differences in the relative responses of the respective mRNAs. The time courses showed the major increase in the initial 6 h; and a decline after 24-36 h. The enhancement of the levels of the mRNAs could be ascribed to transcriptional activation. Since the mitochondrial genome is transcribed as a single polycistronic unit, to account for the > 20-fold differences in the levels of the mitochondrial mRNAs it is necessary to invoke differential stabilities of these mRNAs. The synchronous increase in the expression of both the steroidogenic enzymes and the mitochondrial oxidative phosphorylation system subunits, provides evidence for coregulation of steroidogenic and energy producing capacities of adrenal cells to meet the metabolic needs of steroid hormone production. Suppression of β-actin gene expression may be related to changes in actin polymerization during ACTH-dependent cytoskeletal reorganization. © 1994.
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