Expression and characterization of a fusion protein between the catalytic domain of poly(ADP-ribose) polymerase and the DNA binding domain of the glucocorticoid receptor.
Expression and characterization of a fusion protein between the catalytic domain of poly(ADP-ribose) polymerase and the DNA binding domain of the glucocorticoid receptor.
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聚（ADP-核糖）聚合酶催化结构域和糖皮质激素受体 DNA 结合结构域之间融合蛋白的表达和表征。
DOI：
10.1006/bbrc.1994.2012
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发表时间：
1994
影响因子：
3.1
通讯作者：
Smulson,M
中科院分区：
文献类型：
--
作者：
Rosenthal,D;Hong,T;Cherney,B;Zhang,S;Shima,T;Danielsen,M;Smulson,M
将糖皮质激素受体的DNA结合区与聚腺苷二磷酸核糖聚合酶的催化结构域融合，构建了糖皮质激素受体的融合蛋白。该嵌合蛋白在E.大肠杆菌在真核细胞中，并识别的抗体聚合酶和糖皮质激素受体。类似于聚合酶，嵌合体产生真正的聚（ADP-核糖）聚合物共价结合到蛋白质，并被3-氨基苯甲酰胺抑制。与真正的糖皮质激素受体一样，融合蛋白与含有糖皮质激素反应元件的DNA形成稳定的复合物。在哺乳动物细胞中，融合蛋白显著且特异性地抑制糖皮质激素受体刺激报告构建体的能力。这些结果表明，聚合酶活性可以靶向特定的DNA序列和调节基因表达。
A fusion protein comprising the DNA-binding region of the glucocorticoid receptor and the catalytic domain of poly(ADP-ribose) polymerase was constructed. This chimeric protein was expressed both inE. coliand in eukaryotic cells and was recognized by antibodies to both polymerase and the glucocorticoid receptor. Similar to polymerase, the chimera produced bona fide poly (ADP-ribose) polymers covalently bound to protein and was inhibited by 3-aminobenzamide. Like the authentic glucocorticoid receptor, the fusion protein formed a stable complex with DNA containing the glucocorticoid response element. In mammalian cells, the fusion protein significantly and specifically inhibited the ability of the glucocorticoid receptor to stimulate a reporter construct. These results indicate that polymerase activity can be targeted to specific DNA sequences and modulate gene expression.