Identification of a retinoid/chicken ovalbumin upstream promoter transcription factor response element in the human retinoid X receptor gamma2 gene promoter.

To investigate the mechanisms involved in the transcriptional control of retinoid X receptor (RXR) gene expression, the 5'-flanking region of the human RXRgamma2 isoform was characterized. An imperfect hexamer repeat (gamma retinoid X response element; gammaRXRE) with a single nucleotide spacer (GGTTGAaAGGTCA) was identified immediately upstream of the RXRgamma2 gene transcription start site. Cotransfection studies in CV-1 cells with expression vectors for the retinoid receptors RXRalpha and retinoic acid receptor beta (RARbeta) demonstrated that the gammaRXRE confers retinoid-mediated transcriptional activation with preferential activation by RXR in the presence of its cognate ligand, 9-cis-retinoic acid (RA). Electrophoretic mobility shift assays demonstrated that RXR homodimer binding to gammaRXRE is markedly enhanced by 9-cis-RA, whereas RAR.RXR heterodimer binding is ligand-independent. DNA binding studies and cell cotransfection experiments also demonstrated that the nuclear receptor, chicken ovalbumin upstream promoter transcription factor (COUP-TF), repressed transcription via the gammaRXRE. Cotransfection experiments revealed that COUP-TF and RXRalpha compete at the gammaRXRE to modulate transcription bidirectionally over a wide range. These results demonstrate that the human RXRgamma2 gene promoter contains a novel imperfect repeat element capable of mediating RXR-dependent transcriptional autoactivation and COUP-TF-dependent repression.