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引用次数: 1
摘要
RNA聚合酶II (RNA polymerase II, Pol II)是一种从酵母到人类的12亚基蛋白复合物，是基因表达所必需的。含有Pol II [Pol II (G)]的Gdown1是Pol II的一种特殊形式，具有催化活性，在体外高度依赖于30亚基中介复合物的激活剂和基础依赖功能。在这里，我们首次报道了通过对Pol II (G)完全响应的新型多bac杆状病毒表达系统鉴定和产生15个亚基的人类中介复合物。我们的结果表明，我们合成的完整的30个亚基的中介和15个亚基的重组中介完全恢复了Pol II (G)依赖的转录。此外，我们还发现重组中介体也与Pol II (G)相互作用。这些结果启发我们了解参与选择基因调控的特定Pol II群体是如何被中介复合物激活的。
Reconstitution of Pol II (G) responsive form of the human Mediator complex.
RNA polymerase II (Pol II) is a 12 subunit protein complex from yeast to human that is required for gene expression. Gdown1 containing Pol II [Pol II (G)] is a special form of Pol II that is catalytically inactive and heavily depends on the 30-subunit Mediator complex for its activator and basal dependent function in vitro. Here we report for the first time, the identification and the generation of a 15-subunit human Mediator complex via the novel multibac baculovirus expression system that is fully responsive to Pol II (G). Our results show complete recovery of Pol II (G) dependent transcription both with full 30-subunit Mediator and also with 15-subunit recombinant Mediator that we synthesized. Moreover, we also show that the recombinant Mediator interacts with Pol II (G) as well. These results enlighten us towards understanding how a certain population of Pol II that is involved in selected gene regulation is activated by Mediator complex.
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