大鼠钙结合蛋白D-9k基因5'-侧翼区域中1,25-二羟基维生素D3反应元件的鉴定。

Identification of a 1,25-dihydroxyvitamin D3-response element in the 5'-flanking region of the rat calbindin D-9k gene.

作者信息

Darwish H M, DeLuca H F

机构信息

Department of Biochemistry, University of Wisconsin-Madison 53706.

出版信息

Proc Natl Acad Sci U S A. 1992 Jan 15;89(2):603-7. doi: 10.1073/pnas.89.2.603.

The rat calbindin D-9k gene is transcriptionally regulated by 1,25-dihydroxyvitamin D3 in the intestine. We have examined the 5'-flanking region of this gene and identified a 1,25-dihydroxyvitamin D3-responsive element (DRE) between nucleotides -489 and -445. This element confers 1,25-dihydroxyvitamin D3 responsiveness through its native promoter and the heterologous thymidine kinase promoter, and it contains the sequence GGGTGTCGGAAGCCC, which is homologous to the other previously identified DREs. Incubation of this element with the 1,25-dihydroxyvitamin D3 receptor produced a specific protein-DNA complex, which shifted to a higher molecular weight form upon the addition of a monoclonal antibody specific to the 1,25-dihydroxyvitamin D3 receptor. Therefore, the 5'-flanking region of the rat calbindin D-9k gene contains a DRE that mediates the enhanced expression of this gene by 1,25-dihydroxyvitamin D3 in the intestine.

大鼠钙结合蛋白D-9k基因在肠道中受1,25-二羟基维生素D3的转录调控。我们研究了该基因的5'-侧翼区域，在核苷酸-489至-445之间鉴定出一个1,25-二羟基维生素D3反应元件（DRE）。该元件通过其天然启动子和异源胸苷激酶启动子赋予1,25-二羟基维生素D3反应性，并且它包含序列GGGTGTCGGAAGCCC，该序列与其他先前鉴定的DREs同源。将该元件与1,25-二羟基维生素D3受体一起孵育产生了一种特异性的蛋白质-DNA复合物，在加入针对1,25-二羟基维生素D3受体的单克隆抗体后，该复合物转变为更高分子量的形式。因此，大鼠钙结合蛋白D-9k基因的5'-侧翼区域包含一个DRE，该DRE介导1,25-二羟基维生素D3在肠道中增强该基因的表达。