THE C-FOS CAMP-RESPONSE ELEMENT - REGULATION OF GENE-EXPRESSION BY A BETA-2-ADRENERGIC AGONIST, SERUM AND DNA METHYLATION

The transcription control region of the Proto-oncogene c-fos contains multiple cis-acting elements to which specific trans-acting factors bind. One such well-studied binding motif in the c-fos promoter is the major cyclic AMP response element (CRE) TGACGT located at -62/-57. In this study we investigated the role of this element in gene regulation by beta2-adrenergic/adenylate cyclase signalling and DNA methylation. By transient gene expression assays we were able to show that the c-fos regulatory sequences spanning nucleotides -361 to +13 could mediate gene expression by the beta2-adrenergic agonist isoproterenol and the phosphodiesterase inhibitor theophylline. For isoproterenol however, a stimulating effect was observed in serum-starved cells, while an inhibitory effect was measured in cells supplemented with serum. The gene regulation by the cAMP elevating agents could be due, at least partially, to the major CRE since this isolated motif mediated gene expression by these drugs. Distinct protein-DNA complexes were obtained with nuclear extracts prepared from cells exposed to isoproterenol or/and theophylline under different serum conditions. We further show that DNA methylation of this CRE may also be involved in gene regulation as methylation of the CRE motif strongly reduced the binding of nuclear proteins.