Structural basis of rotavirus RNA chaperone displacement and RNA annealing.
Structural basis of rotavirus RNA chaperone displacement and RNA annealing.
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DOI：
10.1073/pnas.2100198118
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发表时间：
2021-10-12
影响因子：
11.1
通讯作者：
Borodavka A
中科院分区：
文献类型：
--
作者：
Bravo JPK;Bartnik K;Venditti L;Acker J;Gail EH;Colyer A;Davidovich C;Lamb DC;Tuma R;Calabrese AN;Borodavka A
准确的RNA折叠对病毒复制至关重要。轮状病毒是感染人类和动物的病毒。轮状病毒基因组由11个不同的RNA组成，而成功的复制需要将所有11个RNA整合到一个病毒粒子中。RNA伴侣NSP2结合病毒转录本，调节它们之间的相互作用。NSP2必须在碱基对后释放RNA，然后才能包装。利用单分子荧光工具，我们剖析了NSP2的RNA伴侣活性的各个步骤。NSP2-RNA复合体的结构蛋白质组学和冷冻EM研究表明，NSP2通过其带电的C-末端区域调节RNA的展开和释放。病毒RNA伴侣调节的某些方面反映了细菌RNA伴侣所采用的保守的自动调节机制。轮状病毒基因组分布在11个不同的RNA分子中，所有这些分子在病毒组装过程中都必须有选择地被包装起来。这可能是通过RNA伴侣NSP2促进的序列特异性RNA相互作用而发生的。在这里，我们报告了NSP2通过其C-末端区域(CTR)自动调节其伴侣活性，该区域通过限制其螺旋解离活性来促进RNA-RNA相互作用。出乎意料的是，结构蛋白质组学数据显示，CTR并不直接与RNA相互作用，而是加速从NSP2释放RNA。冷冻电子显微镜重建的NSP2-RNA复合体显示，CTR上有一个高度保守的酸性斑块，它倾向于结合的RNA。病毒复制被酸性斑块内的电荷干扰突变所消除，但通过电荷保持突变完全恢复。NSP2和不相关的细菌RNA伴侣Hfq之间的机制相似性表明，加速RNA解离同时促进分子间RNA相互作用可能是RNA伴侣回收的一种普遍策略。
Accurate RNA folding is essential for virus replication. Rotaviruses are viruses infecting humans and animals. Rotavirus genome comprises 11 distinct RNAs, and successful replication requires the incorporation of all 11 RNAs into a virion. The RNA chaperone NSP2 binds viral transcripts, regulating their interactions with each other. NSP2 must release RNAs after they base pair prior to their packaging. Using single-molecule fluorescence tools, we dissected the individual steps of the RNA chaperone activity of NSP2. Structural proteomics and cryo-EM studies of the NSP2–RNA complex revealed that NSP2 regulates RNA unfolding and the release of the RNA using its charged C-terminal region. Some aspects of the viral RNA chaperone regulation mirror the conserved autoregulation mechanisms employed by bacterial RNA chaperones. Rotavirus genomes are distributed between 11 distinct RNA molecules, all of which must be selectively copackaged during virus assembly. This likely occurs through sequence-specific RNA interactions facilitated by the RNA chaperone NSP2. Here, we report that NSP2 autoregulates its chaperone activity through its C-terminal region (CTR) that promotes RNA–RNA interactions by limiting its helix-unwinding activity. Unexpectedly, structural proteomics data revealed that the CTR does not directly interact with RNA, while accelerating RNA release from NSP2. Cryo–electron microscopy reconstructions of an NSP2–RNA complex reveal a highly conserved acidic patch on the CTR, which is poised toward the bound RNA. Virus replication was abrogated by charge-disrupting mutations within the acidic patch but completely restored by charge-preserving mutations. Mechanistic similarities between NSP2 and the unrelated bacterial RNA chaperone Hfq suggest that accelerating RNA dissociation while promoting intermolecular RNA interactions may be a widespread strategy of RNA chaperone recycling.
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影响因子：
5.6
作者：
Carpio, Rodrigo Vasquez-Del;Gonzalez-Nilo, Fernando D.;Patton, John T.
通讯作者：
Patton, John T.
影响因子：
14.9
作者：
Borodavka A;Ault J;Stockley PG;Tuma R
通讯作者：
Tuma R
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
DOI：
10.1146/annurev-virology-092917-043448
发表时间：
2018-01-01
期刊：
ANNUAL REVIEW OF VIROLOGY, VOL 5
影响因子：
--
作者：
Bieniasz, Paul;Telesnitsky, Alice
通讯作者：
Telesnitsky, Alice
影响因子：
15
作者：
Cordes, Thorben;Vogelsang, Jan;Tinnefeld, Philip
通讯作者：
Tinnefeld, Philip