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  • Title: Transcriptional activation of Gs alpha expression by retinoic acid and parathyroid hormone-related protein in F9 teratocarcinoma cells.
    Author: Chan SD, Strewler GJ, Nissenson RA.
    Journal: J Biol Chem; 1990 Nov 25; 265(33):20081-4. PubMed ID: 2122968.
    Abstract:
    Cyclic AMP is known to enhance retinoic acid-induced differentiation of F9 mouse teratocarcinoma cells to parietal endoderm. Recently, we showed that a parathyroid hormone-related protein (PTHrP), by activating adenylate cyclase, can substitute for exogenous cAMP in promoting retinoic acid-induced differentiation of F9 cells. However, the mechanisms by which endogenous cAMP levels are regulated during F9 differentiation are poorly defined. We therefore assessed whether Gs alpha, a subunit of the stimulatory coupling protein of adenylate cyclase, is induced during this process. Treatment of F9 cells with retinoic acid (1 microM) for 5 days resulted in a 20-fold increase in steady-state levels of a 2.0-kilobase Gs alpha mRNA. This was accompanied by an increase in the expression of 52- and 45-kDa Gs alpha polypeptides. Gs alpha mRNA increases within 24 h of exposure to retinoic acid, whereas the expression of alpha 1 (IV) collagen, a marker for F9 differentiation, did not increase until 48 h of treatment. In the presence of retinoic acid, exogenous human PTHrP-(1-34)-amide (20 nM) produced a further 2-fold increase in Gs alpha mRNA. These effects of retinoic acid and PTHrP were exerted largely if not entirely at the level of Gs alpha gene transcription, as assessed by nuclear run-on assay. Bt2cAMP (1 mM) did not reproduce the stimulatory effects of PTHrP on Gs alpha transcription, mRNA, or protein. These data demonstrate that a marked increase in Gs alpha expression accompanies F9 differentiation induced by retinoic acid and PTHrP, and that the regulation is predominantly transcriptional. The resulting increase in adenylate cyclase responsiveness to PTHrP and perhaps other ligands may be a critical component of the differentiation process. The effect of PTHrP on the expression of Gs alpha appears to be mediated by signals other than cAMP.
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