Reference - Detail
RRC ID | 1213 |
---|---|
Author | Kitazawa R, Kitazawa S. |
Title | Vitamin D(3) augments osteoclastogenesis via vitamin D-responsive element of mouse RANKL gene promoter. |
Journal | Biochem Biophys Res Commun |
Abstract |
Receptor activator of NF-kappaB ligand (RANKL) is a membrane-bound signal transducer necessary for the induction and maintenance of osteoclasts. To clarify the molecular mechanism by which 1,25-dihydroxyvitamin D(3) (1,25-(OH)(2)D(3)) augments osteoclasts, we characterized the promoter region of the mouse RANKL gene. Mirroring in vitro osteoclastogenesis demonstrated by a coculture of bone marrow macrophages with ST2 stromal cells, Northern blot, and nuclear run-on analyses showed that 1,25-(OH)(2)D(3) upregulate RANKL gene expression at the transcriptional level. Using a series of deletion mutants of mouse RANKL promoter-luciferase reporter gene constructs, transient transfection studies revealed that the inductive effect of 1,25-(OH)(2)D(3) was abolished when the region up to -723 was deleted. An electrophoretic motility shift assay demonstrated that the VDR-RXRbeta heterodimer bound to AGGTCAGCCTGGTTCA (-937/-922), and VDRE/nuclear protein supershift complexes that bound to anti-VDR and -RXRbeta antibodies were detected in the nuclear extract of 1,25-(OH)(2)D(3)-treated ST2 cells. Furthermore, induction of mutation to the putative VDRE also diminished the inductive effect of 1,25-(OH)(2)D(3). We therefore concluded that mouse RANKL gene is one of the target genes of 1,25-(OH)(2)D(3) containing a functional VDRE in the promoter region. |
Volume | 290(2) |
Pages | 650-5 |
Published | 2002-1-18 |
DOI | 10.1006/bbrc.2001.6251 |
PII | S0006291X01962515 |
PMID | 11785948 |
MeSH | Animals Blotting, Northern Carrier Proteins / genetics* Carrier Proteins / metabolism Cell Differentiation / drug effects Cell Line Cholecalciferol / pharmacology* Coculture Techniques Electrophoresis, Polyacrylamide Gel Gene Expression Regulation / drug effects Gene Expression Regulation / physiology Genes, Reporter Macrophages / cytology Membrane Glycoproteins / genetics* Membrane Glycoproteins / metabolism Mice Mutagenesis, Site-Directed Osteoclasts / cytology Osteoclasts / drug effects* Osteoclasts / metabolism Promoter Regions, Genetic* RANK Ligand Receptor Activator of Nuclear Factor-kappa B Stromal Cells / cytology Stromal Cells / drug effects Stromal Cells / metabolism Transcriptional Activation / drug effects Transfection Up-Regulation / drug effects Vitamin D Response Element / drug effects Vitamin D Response Element / physiology* |
IF | 2.985 |
Times Cited | 73 |
WOS Category | BIOPHYSICS BIOCHEMISTRY & MOLECULAR BIOLOGY |
Resource | |
Human and Animal Cells | ST2(RCB0224) |