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  • Title: Regulation of human corticotropin-releasing hormone gene expression by 3',5'-cyclic adenosine monophosphate in a transformed mouse corticotroph cell line.
    Author: Dorin RI, Takahashi H, Nakai Y, Fukata J, Naitoh Y, Imura H.
    Journal: Mol Endocrinol; 1989 Oct; 3(10):1537-44. PubMed ID: 2558296.
    Abstract:
    In order to characterize potential mechanisms regulating the expression of the human CRH (hCRH) gene, an intact genomic fragment including 5'-flanking sequence of the hCRH gene was stably transfected into the mouse corticotroph AtT20 cell line. The exogenous hCRH gene was expressed at a high frequency with accurate and efficient transcription in transformed cells. Northern blot analysis revealed a single species of CRH mRNA of 1.6 kilobases which was identical in size to human placental CRH mRNA. S1 analysis demonstrated a single cap site in both placenta and transformed AtT20 cells, corresponding to a site 23 base pairs downstream of the TATA box. Treatment with 8-bromo cAMP and phorbol ester resulted in a dose-dependent increase in CRH secretion during a 1-h incubation. Treatment with forskolin or 8-bromo-cAMP also produced a dose-dependent 4- to 10-fold increase in CRH mRNA levels, which was rapid (1 h) and sustained (6, 12, 24, and 48 h). These effects in a well characterized continuous cell culture system demonstrate pretranslational regulation of CRH expression by a cAMP-dependent pathway.
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