Stimulation of oligonucleotide binding of estradiol receptor complexes by accessory proteins.
Stimulation of oligonucleotide binding of estradiol receptor complexes by accessory proteins.
复制标题
辅助蛋白刺激寡核苷酸与雌二醇受体复合物的结合。
DOI：
10.1093/nar/6.12.3859
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发表时间：
1979
影响因子：
14.9
通讯作者：
H. Dickerman
中科院分区：
文献类型：
--
作者：
K. Thanki;T. Beach;A. Bass;H. Dickerman
在使用寡聚(dT)-纤维素层析纯化E2R的过程中，在小鼠肾脏的细胞质中鉴定出了受体辅助因子(RAF)。该因子刺激纯化的 E2R 与寡聚 (dT)、寡聚 (dC) 和寡聚 (dA)-纤维素以及 DNA 纤维素的结合。它是一种热稳定、抗胰蛋白酶的蛋白质，表观分子量在 10 至 30,000 道尔顿之间。尽管结构上不相关，但小牛胸腺组蛋白和蛋清溶菌酶也观察到类似的寡核苷酸结合刺激作用。各个组蛋白，尤其是 H2a、H2B 和 H3，也有利于纯化的 E2R 与寡 (dT)-纤维素的重新结合，而 H1 的效果较差。此外，组蛋白在 4 摄氏度和 12 摄氏度的沉降过程中稳定全感受器。H2b 的 N 端和 C 端半分子是通过溴化氰介导的裂解产生的，并且发现 N 端半分子在结合和全感受器稳定方面复制了母体分子的作用。这些数据表明 E2R 与 DNA 的体内结合可以通过细胞质和细胞核来源的辅助蛋白进行调节。
During purification of E2R using oligo(dT)-cellulose chromatography, a receptor accessory factor (RAF) was identified in the cytosol of mouse kidney. This factor stimulates the binding of purified E2R to oligo(dT)-, oligo(dC)-, and oligo(dA)-cellulose as well as to DNA cellulose. It is a heat-stable, trypsin-resistant protein with an apparent molecular weight of between 10 and 30,000 daltons. Although structurally unrelated, similar stimulation of oligonucleotide binding was seen with calf thymus histones and, to a lesser extent, egg white lysozyme. Individual histones, especially H2a, H2B, and H3, also facilitate rebinding of purified E2R to oligo(dT)-cellulose, while H1 is less effective. Furthermore, histones stabilize the holoreceptor during sedimentation at 4 degrees and 12 degrees C. The N- and C-terminal half molecules of H2b were generated by cyanogen bromide-mediated cleavage and the N-terminal half was found to duplicate the effects of the parent molecule, both in binding and holoreceptor stabilization. These data suggest that the in vivo binding of E2R to DNA can be modulated by accessory proteins of cytosol and nuclear origin.