Title

A central dinucleotide within vitamin D response elements modulates DNA binding and transactivation by the vitamin D receptor in cellular response to natural and synthetic ligands

UMMS Affiliation

Graduate School of Biomedical Sciences; Department of Internal Medicine; Department of Cell Biology

Date

2-9-2002

Document Type

Article

Medical Subject Headings

Animals; Base Sequence; DNA Primers; Electrophoretic Mobility Shift Assay; Humans; Ligands; Osteocalcin; Osteopontin; Protein Binding; RNA, Messenger; Receptors, Calcitriol; Receptors, Retinoic Acid; Retinoid X Receptors; Sialoglycoproteins; *Trans-Activation (Genetics); Transcription Factors; Tumor Cells, Cultured; *Vitamin D Response Element

Disciplines

Life Sciences | Medicine and Health Sciences

Abstract

There is considerable divergence in the sequences of steroid receptor response elements, including the vitamin D response elements (VDREs). Two major VDRE-containing and thus 1,25-dihydroxyvitamin D(3) (1,25-(OH)(2)D(3))-regulated genes are the two non-collagenous, osteoblast-derived bone matrix proteins osteocalcin and osteopontin. We observed a stronger induction of osteopontin than osteocalcin mRNA expression by 1,25-(OH)(2)D(3). Subsequently, we have shown that vitamin D receptor/retinoid X receptor alpha (VDR/RXRalpha) heterodimers bind more tightly to the osteopontin VDRE than to the osteocalcin VDRE. Studies using point mutants revealed that the internal dinucleotide at positions 3 and 4 of the proximal steroid half-element are most important for modulating the strength of receptor binding. In addition, studies with VDRE-driven luciferase reporter gene constructs revealed that the central dinucleotide influences the transactivation potential of VDR/RXRalpha with the same order of magnitude as that observed in the DNA binding studies. The synthetic vitamin D analog KH1060 is a more potent stimulator of transcription and inducer of VDRE binding of VDR/RXR in the presence of nuclear factors isolated from ROS 17/2.8 osteoblast-like cells than the natural ligand 1,25-(OH)(2)D(3). Interestingly, however, KH1060 is comparable or even less potent than 1,25-(OH)(2)D(3) in stimulating VDRE binding of in vitro synthesized VDR/RXRalpha. Thus, the extent of 1,25-(OH)(2)D(3)- and KH1060-dependent binding of VDR/RXRalpha is specified by a central dinucleotide in the VDRE, and the ligand-induced effects on DNA binding are in part controlled by the cellular context of nuclear proteins.

Rights and Permissions

Citation: J Biol Chem. 2002 Apr 26;277(17):14539-46. Epub 2002 Feb 7. Link to article on publisher's site

Related Resources

Link to Article in PubMed

PubMed ID

11834737