Centre for Structural Systems Biology, Heinrich-Pette-Institute, Leibniz Institute for Experimental Virology, University of Hamburg, D-22607 Hamburg, Germany; email:
Division of Structural Biology, Wellcome Centre for Human Genetics, University of Oxford, Oxford OX3 7BN, United Kingdom.
Annu Rev Virol. 2020 Sep 29;7(1):239-262. doi: 10.1146/annurev-virology-021920-115935. Epub 2020 Jul 6.
Viruses are obligatory intracellular parasites that reprogram host cells upon infection to produce viral progeny. Here, we review recent structural insights into virus-host interactions in bacteria, archaea, and eukaryotes unveiled by cellular electron cryo-tomography (cryoET). This advanced three-dimensional imaging technique of vitreous samples in near-native state has matured over the past two decades and proven powerful in revealing molecular mechanisms underlying viral replication. Initial studies were restricted to cell peripheries and typically focused on early infection steps, analyzing surface proteins and viral entry. Recent developments including cryo-thinning techniques, phase-plate imaging, and correlative approaches have been instrumental in also targeting rare events inside infected cells. When combined with advances in dedicated image analyses and processing methods, details of virus assembly and egress at (sub)nanometer resolution were uncovered. Altogether, we provide a historical and technical perspective and discuss future directions and impacts of cryoET for integrative structural cell biology analyses of viruses.
病毒是专性细胞内寄生虫，在感染后会重新编程宿主细胞以产生病毒后代。在这里，我们综述了近年来利用细胞电子冷冻断层成像技术（cryoET）揭示的细菌、古菌和真核生物中病毒-宿主相互作用的结构新见解。这项先进的近自然状态玻璃态样本的三维成像技术在过去二十年中已经成熟，并在揭示病毒复制的分子机制方面证明非常有效。最初的研究仅限于细胞外周，通常集中在早期感染步骤，分析表面蛋白和病毒进入。最近的发展包括 cryo-thinning 技术、相衬成像和相关方法，对于在感染细胞内部靶向罕见事件也非常有帮助。当与专用图像分析和处理方法的进步相结合时，在（亚）纳米分辨率下揭示了病毒组装和出芽的细节。总之，我们提供了历史和技术视角，并讨论了 cryoET 对病毒综合结构细胞生物学分析的未来方向和影响。
