Higher-order structures of the foot-and-mouth disease virus RNA-dependent RNA polymerase required for genome replication.
Higher-order structures of the foot-and-mouth disease virus RNA-dependent RNA polymerase required for genome replication.
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DOI：
10.1038/s42003-021-02989-z
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发表时间：
2022-01-17
影响因子：
5.9
通讯作者：
Stonehouse NJ
中科院分区：
文献类型：
--
作者：
Loundras EA;Streetley J;Herod MR;Thompson R;Harris M;Bhella D;Stonehouse NJ
许多正义RNA病毒的复制发生在细胞内膜相关的隔室内。它们被认为为复制提供了有利的环境，集中了基本的病毒结构和非结构成分，并保护这些成分免受宿主细胞病原体识别和先天免疫反应的影响。然而，这些结构中的分子相互作用和动力学的细节是非常有限的。复制机制的关键组件之一是依赖于RNA的RNA聚合酶RdRp。这种酶已被证明在体外可以形成更高阶的纤维。在这里，使用口蹄疫病毒的RdRp(称为3DPoll)，我们报告了纤维结构，通过冷冻-EM在~7-9 ä分辨率下解决，揭示了柔性组装的多种构象。对高分辨率坐标的拟合导致了潜在分子间相互作用的定义。我们采用了使用亚基因组复制子系统的突变来探索这些相互作用对复制的重要性。我们使用这些数据来提出高阶3Dpol复合体作为动态支架的作用的模型，在其中可以发生RNA复制。Loundras等人。口蹄疫病毒依赖RNA的RNA聚合酶RdRp的纤毛成分报告。他们证明了基于高阶原纤维的相互作用可以创建多个复杂的结构，在这些结构中可以发生RNA复制。
Replication of many positive-sense RNA viruses occurs within intracellular membrane-associated compartments. These are thought to provide a favourable environment for replication to occur, concentrating essential viral structural and nonstructural components, as well as protecting these components from host-cell pathogen recognition and innate immune responses. However, the details of the molecular interactions and dynamics within these structures is very limited. One of the key components of the replication machinery is the RNA-dependent RNA polymerase, RdRp. This enzyme has been shown to form higher-order fibrils in vitro. Here, using the RdRp from foot-and-mouth disease virus (termed 3Dpol), we report fibril structures, solved at ~7-9 Å resolution by cryo-EM, revealing multiple conformations of a flexible assembly. Fitting high-resolution coordinates led to the definition of potential intermolecular interactions. We employed mutagenesis using a sub-genomic replicon system to probe the importance of these interactions for replication. We use these data to propose models for the role of higher-order 3Dpol complexes as a dynamic scaffold within which RNA replication can occur. Loundras et al. report on the fibril components of the RNA-dependent RNA polymerase RdRp from foot-and-mouth disease virus. They demonstrate that higher-order fibril-based interactions create multiple complex structures within which RNA replication can occur.
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影响因子：
5.4
作者：
Herod MR;Ferrer-Orta C;Loundras EA;Ward JC;Verdaguer N;Rowlands DJ;Stonehouse NJ
通讯作者：
Stonehouse NJ
影响因子：
4.8
作者：
Ferrer-Orta, C;Arias, A;Verdaguer, N
通讯作者：
Verdaguer, N
影响因子：
5.4
作者：
Luo, GX;Hamatake, RK;Colonno, RJ
通讯作者：
Colonno, RJ
影响因子：
3.7
作者：
BIENZ, K;EGGER, D;BOSSART, W
通讯作者：
BOSSART, W
影响因子：
5.4
作者：
Chinnaswamy, S.;Murali, A.;Kao, C. C.
通讯作者：
Kao, C. C.