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大鼠肝脏7-8S地塞米松受体与脱氧核糖核酸的结合

Binding of the rat liver 7-8 S dexamethasone receptor to deoxyribonucleic acid.

作者信息

Tymoczko J L, Phillips M M, Vernon S M

出版信息

Arch Biochem Biophys. 1984 Apr;230(1):345-54. doi: 10.1016/0003-9861(84)90117-6.

Abstract

The 7-8 S form of the [3H]dexamethasone (9 alpha-fluoro-11 beta,17,21-trihydroxy-16 alpha-methylpregna-1,4-diene-3, 20-dione) receptor from rat liver cytosol can be converted to the 3-4 S form by RNase treatment or high salt, suggesting a salt-sensitive association between the receptor protein and RNA. In DNA-cellulose column assays, the gradient-purified 3-4 S form bound DNA more efficiently than the 7-8 S form, though the 7-8 S form was also capable of binding to DNA-cellulose to a significant extent. Activated 7-8 S dexamethasone receptor could be released from its association with soluble DNA by treatment with DNase I. Sucrose gradient analysis showed that the released receptor sedimented as the 7-8 S form and was sensitive to RNase treatment, which induced a conversion to the 3-4 S form. Activated RNase-generated 3-4 S receptor again displayed a higher degree of binding to soluble DNA and was recovered in the 3-4 S form following DNase extraction. The fact that the 3-4 S form bound immobilized or soluble DNA more efficiently suggests that the associated RNA of the 7-8 S form interferes directly or indirectly with the receptor association with DNA. The observation that the receptor binds to DNA in its 7-8 S form suggests that the receptor complex is capable of binding RNA and DNA concurrently.

摘要

大鼠肝细胞溶胶中[3H]地塞米松(9α-氟-11β,17,21-三羟基-16α-甲基孕甾-1,4-二烯-3,20-二酮)受体的7-8S形式可通过核糖核酸酶处理或高盐转化为3-4S形式,这表明受体蛋白与RNA之间存在盐敏感关联。在DNA纤维素柱分析中,梯度纯化的3-4S形式比7-8S形式更有效地结合DNA,尽管7-8S形式也能在很大程度上与DNA纤维素结合。活化的7-8S地塞米松受体可通过用脱氧核糖核酸酶I处理而从其与可溶性DNA的结合中释放出来。蔗糖梯度分析表明,释放的受体以7-8S形式沉降,并且对核糖核酸酶处理敏感,核糖核酸酶处理会诱导其转化为3-4S形式。活化的核糖核酸酶产生的3-4S受体再次显示出与可溶性DNA的更高结合程度,并且在脱氧核糖核酸酶提取后以3-4S形式回收。3-4S形式更有效地结合固定化或可溶性DNA这一事实表明,7-8S形式的相关RNA直接或间接干扰受体与DNA的结合。受体以其7-8S形式与DNA结合的观察结果表明,受体复合物能够同时结合RNA和DNA。

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