TY - JOUR T1 - SARS-CoV-2 Assembly and Egress Pathway Revealed by Correlative Multi-modal Multi-scale Cryo-imaging JF - bioRxiv DO - 10.1101/2020.11.05.370239 SP - 2020.11.05.370239 AU - Luiza Mendonça AU - Andrew Howe AU - James B. Gilchrist AU - Dapeng Sun AU - Michael L. Knight AU - Laura C. Zanetti-Domingues AU - Benji Bateman AU - Anna-Sophia Krebs AU - Long Chen AU - Julika Radecke AU - Yuewen Sheng AU - Vivian D. Li AU - Tao Ni AU - Ilias Kounatidis AU - Mohamed A. Koronfel AU - Marta Szynkiewicz AU - Maria Harkiolaki AU - Marisa L. Martin-Fernandez AU - William James AU - Peijun Zhang Y1 - 2020/01/01 UR - http://biorxiv.org/content/early/2020/11/05/2020.11.05.370239.abstract N2 - Since the outbreak of the SARS-CoV-2 pandemic, there have been intense structural studies on purified recombinant viral components and inactivated viruses. However, investigation of the SARS-CoV-2 infection in the native cellular context is scarce, and there is a lack of comprehensive knowledge on SARS-CoV-2 replicative cycle. Understanding the genome replication, assembly and egress of SARS-CoV-2, a multistage process that involves different cellular compartments and the activity of many viral and cellular proteins, is critically important as it bears the means of medical intervention to stop infection. Here, we investigated SARS-CoV-2 replication in Vero cells under the near-native frozen-hydrated condition using a unique correlative multi-modal, multi-scale cryo-imaging approach combining soft X-ray cryo-tomography and serial cryoFIB/SEM volume imaging of the entire SARS-CoV-2 infected cell with cryo-electron tomography (cryoET) of cellular lamellae and cell periphery, as well as structure determination of viral components by subtomogram averaging. Our results reveal at the whole cell level profound cytopathic effects of SARS-CoV-2 infection, exemplified by a large amount of heterogeneous vesicles in the cytoplasm for RNA synthesis and virus assembly, formation of membrane tunnels through which viruses exit, and drastic cytoplasm invasion into nucleus. Furthermore, cryoET of cell lamellae reveals how viral RNAs are transported from double-membrane vesicles where they are synthesized to viral assembly sites; how viral spikes and RNPs assist in virus assembly and budding; and how fully assembled virus particles exit the cell, thus stablishing a model of SARS-CoV-2 genome replication, virus assembly and egress pathways.Competing Interest StatementThe authors have declared no competing interest. ER -