Characterization of the AB (AF-1) region in the muscle-specific retinoid X receptor-gamma: evidence that the AF-1 region functions in a cell-specific manner.

The retinoid X receptors alpha, beta and gamma (RXRs) share a highly conserved 'C' region or DNA binding domain (DBD). The conserved 'DE' region or ligand binding domain (LBD) of the RXRs is functionally complex, mediating dimerization and a ligand-dependent activation function (AF-2). The AB or N-terminal region of the RXRs is poorly conserved and encodes a ligand-independent activation function (AF-1). RXR gamma mRNA is preferentially expressed in skeletal and cardiac muscle, however, cell-specific steroid receptor-mediated trans-activation is a poorly understood phenomenon. We utilized the GAL4 hybrid assay system and have demonstrated that RXR gamma contains two functional domains in the AB and DE regions that activate transcription in a ligand-independent and -dependent manner respectively. The functions of the AB (AF-1) and DE (AF-2) domains were regulated by cAMP-dependent protein kinases, furthermore, the function of AF-2 in the LBD was activated by 8-Br-cAMP, independent of 9-cis-retinoic acid treatment. Deletion analysis demonstrated that the AF-1 of RXR gamma, is located between amino acids 1 and 103 and contained multiple motifs that were targets of cAMP-dependent protein kinases. Transfection analyses in non-muscle and myogenic cells clearly demonstrated that: (i) the AF-1 of RXR gamma functions in a muscle-specific manner and is required for optimal ligand-dependent trans-activation from an RXRE; (ii) RXR gamma trans-activates more efficiently in a myogenic background.