Differences in induction of c-fos transcription by cholera toxin-derived cyclic AMP and Ca2+ signal in astrocytes and 3T3 fibroblasts

The B subunit of cholera toxin, a protein which binds specifically to membrane ganglioside GM1, is known to affect cell growth and differentiation. To investigate the mechanism of these cellular responses at the nuclear level, we used the induction of c-fos in astrocytes and 3T3 fibroblasts as a model. Northern blot analysis showed that treatment with B subunit provokes a rapid and transient expression of c-fos mRNA, independent of a measurable increase in cyclic AMP. The B subunit signal, which is mediated by Ca2+, was compared to cholera toxin and other agents which increase intracellular cyclic AMP levels. In transient transfection assays of astrocytes and fibroblasts, functional analysis of c-fos promoter deletions was used to identify the elements involved in transcriptional activation by B subunit. In astrocytes, the DNA region including the serum response element and the cyclic AMP response element (CRE) are equally required, whereas 3T3 cells require only the CRE for maximal induction. A synergistic effect of signal transduction was mediated by calcium and cyclic AMP on the CRE, being positive in 3T3 cells and negative in astrocytes. Diverse regulatory elements may be thus involved in responses of different cell types to the same extracellular signal. Furthermore, a single regulatory element (CRE) can integrate both calcium and cyclic AMP signals in the control of gene expression.