Structural and mechanistic basis for the inhibition of Escherichia coli RNA polymerase by T7 Gp2.
Structural and mechanistic basis for the inhibition of Escherichia coli RNA polymerase by T7 Gp2.
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DOI：
10.1016/j.molcel.2012.06.013
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发表时间：
2012-09-14
期刊：
影响因子：
16
通讯作者：
Wigneshweraraj, Sivaramesh
中科院分区：
文献类型：
--
作者：
James, Ellen;Liu, Minhao;Sheppard, Carol;Mekler, Vladimir;Camara, Beatriz;Liu, Bing;Simpson, Pete;Cota, Ernesto;Severinov, Konstantin;Matthews, Steve;Wigneshweraraj, Sivaramesh
T7噬菌体编码的小分子蛋白Gp 2是一种非DNA结合转录因子，与大肠杆菌（Escherichia coli，Ec）RNA聚合酶（RNA polymerase，RNAp）β′亚基的钳口结构域相互作用，抑制转录活性启动子复合物（RPo）的形成。在这里，我们描述了Gp 2-Ec β′ jaw结构域复合物的高分辨率溶液结构，并表明Gp 2和DNA竞争结合β′ jaw结构域。我们发现Gp 2有效抑制RPo形成需要氨基末端σ70结构域区域1.1（R1.1），并且Gp 2拮抗RPo形成期间R1.1的强制性运动。我们证明，Gp 2抑制RPo的形成不仅通过空间位阻的RNAp-DNA相互作用，但也通过长距离的拮抗作用RNap启动子的相互作用周围的RNp活性中心，可能发生由于重新定位的R1.1由Gp 2。因此，Gp 2对Ec RNAp的抑制定义了一种以前未表征的机制，通过该机制细菌转录受病毒因子调节。描述了T7转录因子Gp 2-E. coliRNAp β′jaw结构域复合物的结构。Gp 2和DNA竞争β′ jaw结构域上的重叠相互作用表面。Gp 2需要σ70区域1.1才能有效地抑制E. coliRNAp。Gp 2抑制σ70区域1.1的专性置换。
The T7 phage-encoded small protein Gp2 is a non-DNA-binding transcription factor that interacts with the jaw domain of the Escherichia coli (Ec) RNA polymerase (RNAp) β′ subunit and inhibits transcriptionally proficient promoter-complex (RPo) formation. Here, we describe the high-resolution solution structure of the Gp2-Ec β′ jaw domain complex and show that Gp2 and DNA compete for binding to the β′ jaw domain. We reveal that efficient inhibition of RPo formation by Gp2 requires the amino-terminal σ70 domain region 1.1 (R1.1), and that Gp2 antagonizes the obligatory movement of R1.1 during RPo formation. We demonstrate that Gp2 inhibits RPo formation not just by steric occlusion of the RNAp-DNA interaction but also through long-range antagonistic effects on RNAp-promoter interactions around the RNAp active center that likely occur due to repositioning of R1.1 by Gp2. The inhibition of Ec RNAp by Gp2 thus defines a previously uncharacterized mechanism by which bacterial transcription is regulated by a viral factor. ► The structure of the T7 transcription factor Gp2-E. coli RNAp β′jaw domain complex is described ► Gp2 and DNA compete for overlapping interaction surface on the β′ jaw domain ► Gp2 requires σ70 region 1.1 to inhibit the E. coli RNAp efficiently ► Gp2 inhibits the obligatory displacement of σ70 region 1.1
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