Differential transcriptional regulation of the apoAl gene by retinoic acid receptor homo- and heterodimers in yeast

Abstract

Several members of the nuclear receptor superfamily including RXR (retinoid X receptor) bind to a specific retinoic acid response element (site A) of the apoAl promoter. However, transcriptional activation of the apoAl gene by different homo- and heterodimeric forms of RXR or RAR (retinoic acid receptors) cannot be evaluated in mammalian cells, which contain endogenous RXR or RAR. In order to circumvent this limitation, we assessed the DNA-binding activities and transcriptional activation of different homo- and heterodimers of these receptors in yeast. Electrophoretic mobility shift assays (EMSA) demonstrated that yeast | expressed RARα does not bind to site A of the apoAl | promoter, whereas binding of RARβ to site A is ligand-dependent. Both RARα and RARβ form heterodimers with RXRα and bind to site A with high affinity. These DNA-binding studies correlate with the transcriptional data, which indicated that RARβ but not RARα activates transcription from site A in response equally well to 9-cis and all-trans retinoic acids. 9-cis RA is a more potent ligand than all-trans RA to activate transcription via RXR/RAR heterodimers. We conclude that this yeast expression system is a useful tool to elucidate the 'transactivaton code' for apoAl site A via specific combinations of different homo and heterodimeric versions of RXR and RAR.