MOLECULAR-CLONING OF COMPLEMENTARY-DNA ENCODING THE AVIAN RECEPTOR FOR VITAMIN-D
MOLECULAR-CLONING OF COMPLEMENTARY-DNA ENCODING THE AVIAN RECEPTOR FOR VITAMIN-D
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DOI：
10.1126/science.3029866
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发表时间：
1987-03-06
期刊：
影响因子：
56.9
通讯作者：
OMALLEY, BW
中科院分区：
文献类型：
--
作者：
MCDONNELL, DP;MANGELSDORF, DJ;OMALLEY, BW
维生素D3受体是介导活性代谢物1，25-二羟基维生素D3 [1，25（OH）2D 3]的核作用的细胞内蛋白质。使用两种受体特异性单克隆抗体从鸡肠道cDNA表达文库中回收该调节蛋白的互补DNA（cDNA）。从该cDNA推导出的氨基酸序列显示了一个高度保守的富含半胱氨酸的区域，该区域与其他类固醇受体的结构域特征和禽成红细胞增多症病毒的gag-erbA癌基因产物具有同源性。通过与该DNA序列杂交选择的RNA指导免疫可沉淀维生素D3受体的无细胞合成。北方印迹分析的多聚腺苷酸化RNA与这些cDNA探针揭示了两个维生素D受体信使RNA（mRNA）的2.6和3.2的酶在受体的鸡组织和一个主要的交叉杂交受体mRNA种类的4.2的酶在小鼠3 T6成纤维细胞。3 T6细胞预先暴露于1，25（OH）2D 3后，4.2-脱氢酶种类显著增加。这个cDNA可能是迄今为止在真核生物中克隆的最稀有的mRNA，也是第一个描述真实维生素的受体序列。
Vitamin D3 receptors are intracellular proteins that mediate the nuclear action of the active metabolite 1,25-dihydroxyvitamin D3 [1,25(OH)2D3]. Two receptor-specific monoclonal antibodies were used to recover the complementary DNA (cDNA) of this regulatory protein from a chicken intestinal .lambda.gt11 cDNA expression library. The amino acid sequences that were deduced from this cDNA revealed a highly conserved cysteine-rich region that displayed homology with a domain characteristic of other steroid receptors and with the gag-erbA oncogene product of avian erythroblastosis virus. RNA selected via hybridization with this DNA sequence directed the cell-free synthesis of immunoprecipitable vitamin D3 receptor. Northern blot analysis of polyadenylated RNA with these cDNA probes revealed two vitamin D receptor messenger RNAs (mRNAs) of 2.6 and 3.2 kilobases in receptor-containing chicken tissues and a major cross-hybridizing receptor mRNA species of 4.2 kilobases in mouse 3T6 fibroblasts. The 4.2-kilobase species was substantially increased by prior exposure of 3T6 cells to 1,25(OH)2D3. This cDNA represents perhaps the rarest mRNA cloned to date in eukaryotes, as well as the first receptor sequence described for an authentic vitamin.