Direct binding of TFEα opens DNA binding cleft of RNA polymerase.
Direct binding of TFEα opens DNA binding cleft of RNA polymerase.
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DOI：
10.1038/s41467-020-19998-x
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发表时间：
2020-11-30
影响因子：
16.6
通讯作者：
Murakami KS
中科院分区：
文献类型：
--
作者：
Jun SH;Hyun J;Cha JS;Kim H;Bartlett MS;Cho HS;Murakami KS
细胞RNA聚合酶（RNAP） DNA结合裂缝的打开是转录起始的必要条件，但其潜在的分子机制尚不清楚。在这里，我们报道了来自古细菌，柯达热球菌（Tko）的RNAP， RNAP- tfe α二元和RNAP- tfe α-启动子DNA三元复合物的低温电镜结构。这些结构揭示了TFEα桥接RNAP钳和柄结构域以打开DNA结合裂缝。将启动子DNA定位到裂缝中关闭它，同时保持TFEα与RNAP移动模块的相互作用。结构和光交联结果还表明，TFEα扩展翼螺旋结构域中保守的芳香残基与启动子DNA相互作用以稳定转录泡。本研究为TFEα的功能提供了结构基础，并阐明了包括古菌和真核rnap在内的含茎rnap转录起始时DNA结合裂缝打开的机制。在含茎古菌和真核RNA聚合酶（RNAP）系统的转录周期中，箝位构象是如何调控的，目前还不清楚。在这里，作者结合冷冻电镜，x射线晶体学和光交联分析来表征来自kodakarensis热球菌的RNAP， RNAP-TFEα二元和RNAP-TFEα-启动子DNA三元复合物的结构特征，并使他们能够描述转录周期早期一般转录因子TFEα和RNAP的动态构象变化。
Opening of the DNA binding cleft of cellular RNA polymerase (RNAP) is necessary for transcription initiation but the underlying molecular mechanism is not known. Here, we report on the cryo-electron microscopy structures of the RNAP, RNAP-TFEα binary, and RNAP-TFEα-promoter DNA ternary complexes from archaea, Thermococcus kodakarensis (Tko). The structures reveal that TFEα bridges the RNAP clamp and stalk domains to open the DNA binding cleft. Positioning of promoter DNA into the cleft closes it while maintaining the TFEα interactions with the RNAP mobile modules. The structures and photo-crosslinking results also suggest that the conserved aromatic residue in the extended winged-helix domain of TFEα interacts with promoter DNA to stabilize the transcription bubble. This study provides a structural basis for the functions of TFEα and elucidates the mechanism by which the DNA binding cleft is opened during transcription initiation in the stalk-containing RNAPs, including archaeal and eukaryotic RNAPs. How clamp conformation is regulated in the transcription cycle of stalk-containing archaeal and eukaryotic RNA polymerase (RNAP) systems is still not well understood. Here, the authors combine cryo-EM, X-ray crystallography and photo-crosslinking assays to structurally characterise RNAP, the RNAP-TFEα binary and RNAP-TFEα-promoter DNA ternary complexes from the archaea Thermococcus kodakarensis and enables them to describe the dynamic conformational changes of the general transcription factor TFEα and RNAP during the early stage of transcription cycle.
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影响因子：
14.9
作者：
Davis E;Chen J;Leon K;Darst SA;Campbell EA
通讯作者：
Campbell EA
影响因子：
64.5
作者：
Feklistov A;Darst SA
通讯作者：
Darst SA
DOI：
10.1107/s2059798318009324
发表时间：
2018-09-01
期刊：
Acta crystallographica. Section D, Structural biology
影响因子：
--
作者：
Afonine PV;Klaholz BP;Moriarty NW;Poon BK;Sobolev OV;Terwilliger TC;Adams PD;Urzhumtsev A
通讯作者：
Urzhumtsev A
影响因子：
16.6
作者：
Jun SH;Hirata A;Kanai T;Santangelo TJ;Imanaka T;Murakami KS
通讯作者：
Murakami KS
影响因子：
16.8
作者：
Gruenberg, Sebastian;Warfield, Linda;Hahn, Steven
通讯作者：
Hahn, Steven