Cryo-EM Reveals Architectural Diversity in Active Rotavirus Particles
Cryo-EM Reveals Architectural Diversity in Active Rotavirus Particles
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DOI：
10.1016/j.csbj.2019.07.019
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发表时间：
2019-01-01
影响因子：
6
通讯作者：
Kelly, Deborah F.
中科院分区：
文献类型：
--
作者：
Hauser, Mary;Dearnaley, William J.;Kelly, Deborah F.
轮状病毒是一种经过充分研究的 RNA 病毒，可导致儿童严重胃肠炎。在病毒进入过程中，病毒粒子的外层脱落，形成双层颗粒 (DLP)，能够执行病毒转录（即 mRNA 合成）并启动感染。虽然轮状病毒 DLP 的非活性形式已得到详细的结构表征，但有关转录活性 DLP 的信息仍然有限。在这里，我们使用冷冻电子显微镜 (cryo-EM) 和 3D 图像重建来比较转录活性与非活性 DLP 中内部蛋白质成分的结构。我们的研究结果表明，随着 mRNA 合成的展开，转录活性 DLP 获得了内部有序性，而非活性 DLP 则保持动态无序状态。对两个不同对称轴上的病毒蛋白/RNA 成分区域进行分析，以提供运动成分的更全面的视图。总而言之，我们的结果提出了活性 DLP 的新观点，这可能使未来的药理学策略能够以消除轮状病毒转录作为治疗方法。 (C) 2019 年作者。由 Elsevier B.V. 代表计算和结构生物技术研究网络出版。
Rotavirus is a well-studied RNA virus that causes severe gastroenteritis in children. During viral entry, the outer layer of the virion is shed, creating a double-layered particle (DLP) that is competent to perform viral transcription (i.e., mRNA synthesis) and launch infection. While inactive forms of rotavirus DLPs have been structurally characterized in detail, information about the transcriptionally-active DLP remains limited. Here, we used cryo-Electron Microscopy (cryo-EM) and 3D image reconstructions to compare the structures of internal protein components in transcriptionally-active versus inactive DLPs. Our findings showed that transcriptionally-active DLPs gained internal order as mRNA synthesis unfolded, while inactive DLPs remained dynamically disordered. Regions of viral protein/RNA constituents were analyzed across two different axes of symmetry to provide a more comprehensive view of moving components. Taken together, our results bring forth a new view of active DLPs, which may enable future pharmacological strategies aimed at obliterating rotavirus transcription as a therapeutic approach. (C) 2019 The Authors. Published by Elsevier B.V. on behalf of Research Network of Computational and Structural Biotechnology.