Vitamin D3 augments osteoclastogenesis via vitamin D-responsive element of mouse RANKL gene promoter
Vitamin D3 augments osteoclastogenesis via vitamin D-responsive element of mouse RANKL gene promoter
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DOI：
10.1006/bbrc.2001.6251
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发表时间：
2002-01-18
影响因子：
3.1
通讯作者：
Kitazawa, S
中科院分区：
文献类型：
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作者：
Kitazawa, R;Kitazawa, S
NF-B-kappa配体受体激活因子（Receptor activator of NF-B-kappa ligand， RANKL）是一种诱导和维持破骨细胞所需的膜结合信号换能器。为了阐明1,25-二羟基维生素D-3 (1,25-(OH)(2)D-3)增强破骨细胞的分子机制，我们对小鼠RANKL基因的启动子区域进行了表征。骨髓巨噬细胞与ST2基质细胞共培养，Northern blot和核运行分析表明，1,25-(OH)(2)D-3在转录水平上调RANKL基因的表达。利用小鼠RANKL启动子-荧光素酶报告基因构建的一系列缺失突变体，瞬时转染研究发现，当缺失-723以下区域时，125 -(OH)(2)D-3的诱导作用被消除。电泳运动位移实验表明，在1,25-(OH)(2) d -3处理的ST2细胞核提取物中检测到与（AGGTCA）结合的VDR-RXRbeta异源二聚体（-937/-922）下的GCC (TGGTTCA)，以及与抗vdr和- rxrbeta抗体结合的VDRE/核蛋白超位移复合物。此外，诱导突变的假设VDRE也削弱了1,25-(OH)(2)D-3的诱导作用。因此，我们得出结论，小鼠RANKL基因是1,25-(OH)(2)D-3的靶基因之一，该基因在启动子区域含有功能性VDRE。(C) 2002 Elsevier Science。
Receptor activator of NF-B-kappa 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. Am electrophoretic motility shift assay demonstrated that the VDR-RXRbeta heterodimer bound to (AGGTCA) under bar GCC (TGGTTCA) under bar (-937/-922), and VDRE/nuclear protein super-shift 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. (C) 2002 Elsevier Science.