Tumor necrosis factor activates a nuclear inhibitor of vitamin D and retinoid-X receptors.

Tumor necrosis factor-alpha (TNF) is an important contributor to the pathophysiology of bone loss in osteoporosis. Previous work has revealed that TNF inhibits 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) action. We have shown that TNF decreases binding of the vitamin D receptor (VDR) and its heterodimeric partner, the retinoid-x receptor (RXR), to the vitamin D response element (VDRE) of the osteocalcin gene. Here we test the hypothesis that TNF induces a nuclear inhibitor of RXR/VDR binding to DNA and that this inhibitor can have independent effects on RXR. The effect of TNF on RXR and VDR binding to their cognate response elements and stimulation of transcription was studied in VDR deficient CV-1 and COS-7 cells. In CV-1 cells transfected with VDR and RXR expression vectors, TNF-alpha inhibited 1,25(OH)2D3 stimulated transcription of a VDRE-CAT reporter and 9-cis-retinoic acid (9cRA) stimulated transcription of an RXRE-CAT reporter. Inhibition of transcription was associated with decreased binding of VDR and RXR to their cognate response elements. To determine if TNF-alpha induced a nuclear inhibitor of VDR and RXR binding to DNA, nuclear extract was isolated from TNF treated receptor deficient COS cells and mixed with nuclear extract from ligand treated receptor replete COS cells. Receptor binding to DNA was inhibited by the extract from TNF treated COS-7 cells. The inhibitory activity rapidly appeared in nuclear extracts following TNF stimulation. We conclude that TNF activates a nuclear inhibitor of VDR and RXR.