Stability and sequence-specific DNA binding of activation-labile mutants of the human glucocorticoid receptor.
Stability and sequence-specific DNA binding of activation-labile mutants of the human glucocorticoid receptor.
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人类糖皮质激素受体激活不稳定突变体的稳定性和序列特异性 DNA 结合。
DOI：
10.1021/bi00110a017
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发表时间：
1991
期刊：
影响因子：
2.9
通讯作者：
Harmon,JM
中科院分区：
文献类型：
--
作者：
Elsasser,MS;Eisen,LP;Riegel,AT;Harmon,JM
药理学系，健康科学统一服务大学，Bethesda，马里兰州20814-4799，和药理学系，乔治敦大学，3900 Reservoir Road，NW，华盛顿，DC 20007接收日期：1991年4月3日;修订版手册接收日期：1991年7月25日摘要：来自糖皮质激素抗性突变体3R 7的激活不稳定（act 1）人糖皮质激素受体（hGR）的稳定性和DNA结合特性。6TG。4人被调查。这些受体能够以正常的亲和力和特异性可逆地结合结合配体，但在试图活化为DNA结合形式期间变得不稳定[Harmon et al.（1984）J. Steroid Biochem. 21，227-236]。亲和标记和免疫化学分析表明，act 1受体不优先蛋白水解过程中试图激活。此外，结合小牛胸腺DNA的分析表明，在失去配体后，act 1受体保留了非特异性结合DNA的能力。使用含有多个GRE序列的370 bp MMTV启动子片段和缺乏GRE序列的上游342 bp片段来评估act 1 hGR与特定DNA序列的结合。用~（32）P末端标记的片段与hGR-DNA复合物孵育后的免疫吸附实验表明，正常和act 1 hGR均以激活依赖的方式选择性地与含有GR的片段结合。正常hGR和act'hGR的结合可以用含有完美回文GRE的合成寡核苷酸阻断，但不能用GRE被ERE取代的寡核苷酸阻断。在不存在配体的情况下，或在与糖皮质激素拮抗剂RU 38486孵育后，正常和actx hGR激活获得类似的结果。这些结果表明，hGR的序列特异性结合不需要结合配体的存在，并表明配体在神经应答基因的反式激活中的作用。类固醇激素受体调节生殖应答基因表达的能力严格依赖于结合配体的存在，以及类固醇-受体复合物活化成能够与序列特异性DNA结合的形式（Evans，1988; Beato，1989; Carson-Jurica等，1990年）。蛋白水解受体片段的分析和定点诱变将糖皮质激素受体（GR）1的配体结合结构域定位于类固醇结合蛋白的羧基末端三分之一（Carl-stedt-杜克等人，1987; Giguere等人，1986; Danielsen等人，
Department of Pharmacology, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814-4799, and Department of Pharmacology, Georgetown University, 3900 Reservoir Road, NW, Washington, DC 20007 Received April 3, 1991; Revised Manuscript Received July 25, 1991 abstract: The stability and DNA-binding properties of activation-labile (act1) human glucocorticoid receptors (hGRs) from the glucocorticoid-resistant mutant 3R7. 6TG. 4 were investigated. These receptors are able to bind reversibly associating ligands with normal affinity and specificity, but become unstable during attempted activation to the DNA binding form [Harmon et al.(1984) J. Steroid Biochem. 21, 227-236]. Affinity labeling and immunochemical analysis demonstrated that act1 receptors are not preferentially proteolyzed during attempted activation. In addition, analysis of binding to calf thymus DNA showed that after loss of ligand, act1 receptors retain the ability to bind to DNA nonspecifically. A 370 bp MMTV promoter fragment containing multiple GREs and an upstream 342 bp fragment lacking GRE sequences were used to assess the binding of act1 hGR to specific DNA sequences. Immunoadsorption of hGR-DNA complexes after incubation with 32P-end-labeled fragments showed that both normal and act1 hGR bound selectivelyto the GRE-containing fragment in an activation-dependent manner. Binding of both normal and act'hGRs could be blocked with a synthetic oligonucleotide containing a perfect palindromic GRE, but not with an oligonucleotide in which the GRE was replaced by an ERE. Analogous results were obtained for normal and actx hGR activated in the absence of ligand, or after incubation with the gluco-corticoid antagonist RU 38486. These results suggest that sequence-specific binding of the hGR does not require the presence of bound ligand and suggest a role for the ligand in trans-activation of hormonally responsive genes. e ability of steroid hormone receptors to modulate the expression of hormonally responsive genes is strictly dependent upon the presence of bound ligand, and activation of the steroid-receptor complexto a form capable of sequence-spe-cific DNA binding (Evans, 1988; Beato, 1989; Carson-Jurica et al., 1990). Analysis of proteolytic receptor fragments and site-directed mutagenesishave mapped the ligand binding domain of the glucocorticoid receptor (GR) 1to the carbox-yl-terminal one-third of the steroidbinding protein (Carl-stedt-Duke et al., 1987; Giguere et al., 1986; Danielsen et al.,
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影响因子：
64.8
作者：
PICARD, D;KHURSHEED, B;YAMAMOTO, KR
通讯作者：
YAMAMOTO, KR
无细胞系统中雌激素受体依赖性转录机制
影响因子：
5.3
作者：
J. F. Elliston;S. Fawell;L. Klein;S. Tsai;M. Tsai;M. Parker;B. O’Malley
通讯作者：
B. O’Malley
类固醇抗糖皮质激素在淋巴细胞中的作用机制。
DOI：
10.1016/s0021-9258(19)39871-0
发表时间：
1990
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
B. Segnitz;U. Gehring
通讯作者：
U. Gehring
激素的结合加速了糖皮质激素和黄体酮受体与 DNA 结合的动力学。
DOI：
--
发表时间：
1989
影响因子：
11.1
作者：
M. Schauer;G. Chalepakis;T. Willmann;M. Beato
通讯作者：
M. Beato
人淋巴样细胞系 CEM-C7 的钼酸盐敏感和钼酸盐抗性活化不稳定糖皮质激素受体突变体。
DOI：
10.1016/0022-4731(84)90274-7
发表时间：
1984
期刊：
Journal of steroid biochemistry
影响因子：
--
作者：
Harmon,JM;Schmidt,TJ;Thompson,EB
通讯作者：
Thompson,EB