GENETIC COMPLEMENTATION OF A GLUCOCORTICOID RECEPTOR DEFICIENCY BY EXPRESSION OF CLONED RECEPTOR CDNA
GENETIC COMPLEMENTATION OF A GLUCOCORTICOID RECEPTOR DEFICIENCY BY EXPRESSION OF CLONED RECEPTOR CDNA
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DOI：
10.1016/0092-8674(86)90659-8
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发表时间：
1986-08-01
期刊：
影响因子：
64.5
通讯作者：
YAMAMOTO, KR
中科院分区：
文献类型：
--
作者：
MIESFELD, R;RUSCONI, S;YAMAMOTO, KR
我们分离并测序了6.3 kb编码大鼠糖皮质激素受体的cDNA，糖皮质激素受体是一种结合并激活一类激素依赖性转录增强子的蛋白质。含有受体的细胞产生。simeq的受体mrna。6.5 KB和.simeq。4.8 KB，仅在它们的3”非翻译区域有所不同；转录本5”部分的795个氨基酸残基的开放阅读框。编码区在体外、瞬时转染和受体缺陷细胞系的稳定转染中表达。蛋白质产物在沉淀和电泳迁移率、抗体反应性、激素和DNA结合方面与真正的受体难以区分。此外，克隆的受体蛋白激活其相应的增强子，使受体缺陷细胞恢复受调节增强的全部能力。
We isolated and sequenced 6.3 kb of cDNA encoding the rat glucocorticoid receptor, a protein that binds and activates a class of hormone-dependent transcriptional enhancers. Receptor-containing cells produce receptor mRNAs of .simeq. 6.5 kb and .simeq. 4.8 kb that differ only in their 3'' nontranslated regions; an open reading frame of 795 amino acids residues within the 5'' portion of the transcripts. The coding region was expressed in vitro, in transient transfections, and in stable transfectants of a receptor-deficient cell line. The protein products are indistinguishable from bona fide receptor with respect to sedimentation and electrophoretic mobility, antibody reactivity, and hormone and DNA binding. Moreover the cloned receptor protein activates its corresponding enhancers, restoring to the receptor-deficient cells the full capacity for regulated enhancement.