Purified glucocorticoid receptors bind selectively in vitro to a cloned DNA fragment whose transcription is regulated by glucocorticoids in vivo.
Purified glucocorticoid receptors bind selectively in vitro to a cloned DNA fragment whose transcription is regulated by glucocorticoids in vivo.
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纯化的糖皮质激素受体在体外选择性地与克隆的 DNA 片段结合，该片段的转录在体内受到糖皮质激素的调节。
DOI：
10.1073/pnas.78.11.6628
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发表时间：
1981
影响因子：
11.1
通讯作者：
Yamamoto,KR
中科院分区：
文献类型：
--
作者：
Payvar,F;Wrange,O;Carlstedt-Duke,J;Okret,S;Gustafsson,JA;Yamamoto,KR
从大鼠肝脏提取物中纯化至40-60%均一性的活化糖皮质激素受体蛋白，在体外选择性结合小鼠乳腺肿瘤病毒（MTV）DNA的克隆片段。测试的DNA片段含有完整MTV DNA中存在的序列的大约一半，并且当它被引入含有受体的细胞的基因组中时，它的转录速率与完整病毒元件的转录速率一样，受到糖皮质激素的强烈刺激。相反，该受体不能选择性地结合来自大肠杆菌的DNA限制性片段。coli质粒pBR 322和RSF 2124或来自λ和T4噬菌体。定位MTV DNA中负责选择性结合的区域的初步实验迄今已揭示了一个不能结合受体的亚片段和一个远离正常RNA转录物5'末端的下游定位的选择性结合亚片段。这些研究与类固醇受体可能通过识别受调控基因内或附近的特定DNA序列来调节转录速率的观点一致。
Activated glucocorticoid receptor protein, purified to 40-60% homogeneity from rat liver extracts, binds selectively in vitro to a cloned fragment of murine mammary tumor virus (MTV) DNA. The DNA fragment tested contains about half of the sequences present in intact MTV DNA, and its rate of transcription, like that of the intact viral element, is strongly stimulated by glucocorticoids when it is introduced into the genome of a receptor-containing cell. In contrast, the receptor fails to bind selectively to DNA restriction fragments from E. coli plasmids pBR322 and RSF2124 or from bacteriophages lambda and T4. Preliminary experiments to localize regions within MTV DNA responsible for selective binding have revealed thus far one subfragment that fails to bind the receptor and one selectively bound subfragment that maps far downstream from the 5' terminus of the normal RNA transcript. These studies are consistent with the notion that steroid receptors may modulate rates of transcription by recognizing specific DNA sequences within or near the regulated genes.