Structural visualization of RNA polymerase III transcription machineries.

Structural visualization of RNA polymerase III transcription machineries.
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DOI:
10.1038/s41421-018-0044-z
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发表时间:
2018
期刊:
影响因子:
33.5
通讯作者:
He Y
He Y
中科院分区:
生物学1区
文献类型:
--
作者:
Han Y;Yan C;Fishbain S;Ivanov I;He Y

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RNA聚合酶III (Pol III)转录起始需要转录因子IIIB (TFIIIB)的作用,并受到高度调控。在这里,我们使用单粒子低温电子显微镜(cryo-EM)确定了Pol III预引发配合物(PICs)的结构。我们使用模拟各种功能状态的核酸支架观察到稳定的Pol III-TFIIIB复合物,其中TFIIIB紧密环绕上游启动子DNA。TFIIIB和Pol III之间存在复杂的相互作用,稳定了Pol III的C34亚基在活性位点间隙上的翼状螺旋结构域。Pol III PIC的架构更类似于Pol II PIC而不是Pol I PIC。此外,我们还利用延伸复合物(EC)支架获得了与TFIIIB复合物中的Pol III的三维重建,揭示了转录重新启动之前Pol III的促进再循环的机制。
RNA polymerase III (Pol III) transcription initiation requires the action of the transcription factor IIIB (TFIIIB) and is highly regulated. Here, we determine the structures of Pol III pre-initiation complexes (PICs) using single particle cryo-electron microscopy (cryo-EM). We observe stable Pol III–TFIIIB complexes using nucleic acid scaffolds mimicking various functional states, in which TFIIIB tightly encircles the upstream promoter DNA. There is an intricate interaction between TFIIIB and Pol III, which stabilizes the winged-helix domains of the C34 subunit of Pol III over the active site cleft. The architecture of Pol III PIC more resembles that of the Pol II PIC than the Pol I PIC. In addition, we also obtain a 3D reconstruction of Pol III in complex with TFIIIB using the elongation complex (EC) scaffold, shedding light on the mechanism of facilitated recycling of Pol III prior to transcription re-initiation.
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