Vitamin D-mediated regulation of CYP21A2 transcription - A novel mechanism for vitamin D action

Background: 1 alpha,25-Dihydroxyvitamin D-3 has recently been reported to decrease expression and activity of CYP21A2. In this paper, we have studied the mechanisms for the 1 alpha,25-dihydroxyvitamin D-3-mediated effect on CYP21A2 transcriptional rate. Methods: We have studied the effects of 1 alpha,25-dihydroxyvitamin D-3 using luciferase reporter constructs containing different lengths of the CYP21A2 promoter. These constructs were transfected into cell lines derived from human and mouse adrenal cortex. The mechanism for the effects of vitamin Don the CYP21A2 promoter was studied using chromatin immunoprecipitation assay, mutagenesis and gene silencing by siRNA. Results: 1 alpha,25-Dihydroxyvitamin D-3 was found to alter the promoter activity via a VDR-mediated mechanism, including the comodulators VDR interacting repressor (VDIR) and Williams syndrome transcription factor (WSTF). The involvement of comodulator VDIR was confirmed by gene silencing. We identified a vitamin D response element in the CYP21A2 promoter. Interaction between this novel response element and VDR, WSTF and VDIR was shown by chromatin immunoprecipitation assay. When this sequence was deleted, the effect of 1 alpha,25-dihydroxyvitamin D-3 was abolished, indicating that this sequence in the CYP21A2 promoter functions as a vitamin D response element. Interestingly, an altered balance between nuclear receptors and comodulators reversed the suppressing effect of vitamin D to a stimulatory effect. General significance: This paper reports data important for the understanding of the mechanisms for vitamin D-mediated suppression of gene expression as well as for the vitamin D-mediated effects on CYP21A2. We report a novel mechanism for effects of 1 alpha,25-dihydroxyvitamin D-3. (c) 2012 Elsevier B.V. All rights reserved.