The positions of TFIIF and TFIIE in the RNA polymerase II transcription preinitiation complex
The positions of TFIIF and TFIIE in the RNA polymerase II transcription preinitiation complex
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DOI：
10.1038/nsmb1272
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发表时间：
2007-08-01
影响因子：
16.8
通讯作者：
Hahn, Steven
中科院分区：
文献类型：
--
作者：
Chen, Hung-Ta;Warfield, Linda;Hahn, Steven
我们将非天然光反应性氨基酸对苯甲酰基-L-苯丙氨酸 (Bpa) 纳入 Rpb1 和 Rpb2 亚基形成的中央裂口周围的 RNA 聚合酶 II (Pol II) 表面。预引发复合物 (PIC) 与这些 Pol II 衍生物的光交联和羟基自由基裂解测定表明，TFIIF 二聚化结构域与 Rpb9 附近的 Rpb2 叶和突出结构域相互作用，而 TFIIE 与 Pol II 中央裂口另一侧的 Rpb1 钳结构域交联。 Rpb2 叶和突出结构域的突变改变了 Pol II-TFIIF 结合和转录起始位点，这是与 TFIIF、Rpb9 和 TFIIB 突变相关的表型。与之前的生化和结构研究一起，这些发现阐明了 PIC 的结构组织以及参与转录起始位点选择的蛋白质-蛋白质相互作用网络。
We incorporated the non-natural photoreactive amino acid p-benzoyl-L-phenylalanine (Bpa) into the RNA polymerase II (Pol II) surface surrounding the central cleft formed by the Rpb1 and Rpb2 subunits. Photo-cross-linking of preinitiation complexes (PICs) with these Pol II derivatives and hydroxyl-radical cleavage assays revealed that the TFIIF dimerization domain interacts with the Rpb2 lobe and protrusion domains adjacent to Rpb9, while TFIIE cross-links to the Rpb1 clamp domain on the opposite side of the Pol II central cleft. Mutations in the Rpb2 lobe and protrusion domains alter both Pol II-TFIIF binding and the transcription start site, a phenotype associated with mutations in TFIIF, Rpb9 and TFIIB. Together with previous biochemical and structural studies, these findings illuminate the structural organization of the PIC and the network of protein-protein interactions involved in transcription start site selection.