Interplay of positive and negative effectors in function of the C-terminal repeat domain of RNA polymerase II.
Interplay of positive and negative effectors in function of the C-terminal repeat domain of RNA polymerase II.
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RNA 聚合酶 II C 端重复结构域功能中正负效应子的相互作用。
DOI：
10.1073/pnas.91.6.2362
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发表时间：
1994
影响因子：
11.1
通讯作者：
Kornberg,RD
中科院分区：
文献类型：
--
作者：
Li,Y;Kornberg,RD
RNA聚合酶II缺乏一个C-末端结构域（CTD）是活跃的转录与纯化的蛋白质从酵母，但未能支持在酵母提取物中的转录。CTD依赖性可以通过添加来自提取物的两个级分在纯化的系统中重建。抑制部分废除野生型和CTD-少RNA聚合酶的转录;刺激部分恢复野生型聚合酶的活性，但对CTD-少酶的影响要小得多。使用激酶抑制剂可防止CTD被RNA聚合酶II起始因子B磷酸化，获得了平行结果。激酶抑制剂废除野生型聚合酶在酵母提取物中的转录，但在纯化的系统中没有显着的效果。提取物中转录对CTD和激酶作用的要求表明CTD磷酸化参与对抗提取物中的负效应物。除了CTD的磷酸化之外，因子B必须发挥作用，因为在纯化的系统中，它仍然是用缺乏CTD的聚合酶转录所必需的。
RNA polymerase II lacking a C-terminal domain (CTD) was active in transcription with purified proteins from yeast but failed to support transcription in a yeast extract. CTD dependence could be reconstituted in the purified system by addition of two fractions from the extract. An inhibitory fraction abolished transcription by both wild-type and CTD-less RNA polymerases; a stimulatory fraction restored activity of the wild-type polymerase but had a much lesser effect on the CTD-less enzyme. Parallel results were obtained with the use of a kinase inhibitor that prevents phosphorylation of the CTD by RNA polymerase II initiation factor b. The kinase inhibitor abolished transcription by wild-type polymerase in yeast extract but had no significant effect in the purified system. The requirement for both the CTD and kinase action for transcription in an extract indicates that CTD phosphorylation is involved in opposing the negative effector in the extract. Factor b must play a role(s) in addition to phosphorylation of the CTD because it was still required for transcription with polymerase lacking a CTD in the purified system.