The genes of the coactivator TIF2 and the corepressor SMRT are primary 1α,25(OH) 2D 3 targets

Abstract

The complex of the receptor for the hormone 1α,25-dihydroxyvitamin D 3 (1α,25(OH) 2D 3), Vitamin D 3 receptor (VDR), the retinoid X receptor (RXR) and a 1α,25(OH) 2D 3 response element (VDRE) is considered to be the molecular switch for nuclear 1α,25(OH) 2D 3 signaling. In the presence of ligand the VDR–RXR complex interacts with coactivator (CoA) proteins that in turn contact components of the basal transcriptional machinery resulting in an enhanced transcription of 1α,25(OH) 2D 3 target genes. In the absence of ligand the VDR remains bound to the DNA and interacts with corepressor (CoR) proteins that are involved in gene silencing activity. We treated MCF-7 breast cancer cells with 1α,25(OH) 2D 3 for increasing amounts of time, extracted mRNA and screened by real-time PCR the members of the p160 CoA and NCoR CoR families. We find that of the p160 coactivators, only TIF2 was responsive to 1α,25(OH) 2D 3. Similarly SMRT but not NCoR1 gene transcription was sensitive to 1α,25(OH) 2D 3 treatment. In silico analysis revealed that both TIF2 and SMRT promoters have substantial numbers of VDREs compared to the promoters of the other family members. These VDREs are formed by direct repeats of the core binding motif RGKTCA with a three nucleotide spacing (DR3). We suggest that some or all of these DR3-type VDREs are responsible for the observed responsiveness of TIF2 and SMRT to 1α,25(OH) 2D 3.