A novel RNA pol II CTD interaction site on the mRNA capping enzyme is essential for its allosteric activation.
A novel RNA pol II CTD interaction site on the mRNA capping enzyme is essential for its allosteric activation.
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DOI：
10.1093/nar/gkab130
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发表时间：
2021-04-06
影响因子：
14.9
通讯作者：
Pisliakov AV
中科院分区：
文献类型：
--
作者：
Bage MG;Almohammed R;Cowling VH;Pisliakov AV
mRNA加帽酶（CE/RNGTT）募集到转录位点对于5 'mRNA帽的形成至关重要，这反过来又确保了真核细胞中mRNA的高效转录、剪接、多聚腺苷酸化、核输出和翻译。CE GT酶由RNA聚合酶II的丝氨酸-5磷酸化羧基末端结构域（CTD）募集和激活。通过使用分子动力学模拟和增强的采样技术，我们提供了一个系统的和详细的表征的人CE-CTD接口，描述的CTD磷酸化状态，长度和方向上的这种相互作用的影响。我们的计算分析确定了新的CTD相互作用的网站上的人CE GTase表面和量化他们的相对贡献CTD结合。我们还确定，第一次，变构连接之间的CE GTase活性位点和CTD结合位点，使我们能够提出一种机制，变构激活。通过结合和活性测定，我们验证了新型CTD结合位点，并表明CDS 2位点对于CE GTase活性刺激至关重要。在不同的真核生物类群的CE-CTD复合物的共晶结构的新网站的比较表明，这个接口是相当保守的比以前的结构已经表明。
Recruitment of the mRNA capping enzyme (CE/RNGTT) to the site of transcription is essential for the formation of the 5′ mRNA cap, which in turn ensures efficient transcription, splicing, polyadenylation, nuclear export and translation of mRNA in eukaryotic cells. The CE GTase is recruited and activated by the Serine-5 phosphorylated carboxyl-terminal domain (CTD) of RNA polymerase II. Through the use of molecular dynamics simulations and enhanced sampling techniques, we provide a systematic and detailed characterization of the human CE–CTD interface, describing the effect of the CTD phosphorylation state, length and orientation on this interaction. Our computational analyses identify novel CTD interaction sites on the human CE GTase surface and quantify their relative contributions to CTD binding. We also identify, for the first time, allosteric connections between the CE GTase active site and the CTD binding sites, allowing us to propose a mechanism for allosteric activation. Through binding and activity assays we validate the novel CTD binding sites and show that the CDS2 site is essential for CE GTase activity stimulation. Comparison of the novel sites with cocrystal structures of the CE–CTD complex in different eukaryotic taxa reveals that this interface is considerably more conserved than previous structures have indicated.
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影响因子：
15
作者：
Swift RV;McCammon JA
通讯作者：
McCammon JA
DOI：
10.1073/pnas.1106610108
发表时间：
2011-06-21
影响因子：
11.1
作者：
Chu, Chun;Das, Kalyan;Shatkin, Aaron J.
通讯作者：
Shatkin, Aaron J.
影响因子：
4.6
作者：
Bharati, Akhilendra Pratap;Singh, Neha;Akhtar, Md. Sohail
通讯作者：
Akhtar, Md. Sohail
影响因子：
64.5
作者：
Cheng, Hong;Dufu, Kobina;Reed, Robin
通讯作者：
Reed, Robin
影响因子：
4.3
作者：
Alam, Nawsad;Goldstein, Oriel;Schueler-Furman, Ora
通讯作者：
Schueler-Furman, Ora