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

Gert Jan C M Van Den Bemd, Mila Jhamai, Ada Staal, Andre J van Wijnen, Jane B. Lian, Gary S. Stein, Huibert A P Pols, Johannes P T M Van Leeuwen

Research output: Contribution to journalArticle

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Abstract

There is considerable divergence in the sequences of steroid receptor response elements, including the vitamin D response elements (VDREs). Two major VDREcontaining and thus 1,25-dihydroxyvitamin D3 (1,25- (OH)2D3)-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)2D3. Subsequently, we have shown that vitamin D receptor/retinoid X receptor α (VDR/RXCRα) 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/ RXRα 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)2D3. Interestingly, however, KH1060 is comparable or even less potent than 1,25-(OH)2D3 in stimulating VDRE binding of in vitro synthesized VDR/RXRα. Thus, the extent of 1,25-(OH)2D3- and KH1060-dependent binding of VDR/RXRα 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.

Original languageEnglish (US)
Pages (from-to)14539-14546
Number of pages8
JournalJournal of Biological Chemistry
Volume277
Issue number17
DOIs
StatePublished - Apr 26 2002
Externally publishedYes

Fingerprint

Vitamin D Response Element
Calcitriol Receptors
Transcriptional Activation
Ligands
DNA
Osteopontin
Osteocalcin
Osteoblasts
Genes
Retinoid X Receptors
Bone Matrix
Calcitriol
Steroid Receptors
Response Elements
Transcription
Nuclear Proteins
Luciferases
Reporter Genes
Vitamin D
Bone

ASJC Scopus subject areas

  • Biochemistry

Cite this

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. / Van Den Bemd, Gert Jan C M; Jhamai, Mila; Staal, Ada; van Wijnen, Andre J; Lian, Jane B.; Stein, Gary S.; Pols, Huibert A P; Van Leeuwen, Johannes P T M.

In: Journal of Biological Chemistry, Vol. 277, No. 17, 26.04.2002, p. 14539-14546.

Research output: Contribution to journalArticle

Van Den Bemd, Gert Jan C M ; Jhamai, Mila ; Staal, Ada ; van Wijnen, Andre J ; Lian, Jane B. ; Stein, Gary S. ; Pols, Huibert A P ; Van Leeuwen, Johannes P T M. / 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. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 17. pp. 14539-14546.
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T1 - 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

AU - Van Den Bemd, Gert Jan C M

AU - Jhamai, Mila

AU - Staal, Ada

AU - van Wijnen, Andre J

AU - Lian, Jane B.

AU - Stein, Gary S.

AU - Pols, Huibert A P

AU - Van Leeuwen, Johannes P T M

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