Correlative multi-scale cryo-imaging unveils SARS-CoV-2 assembly and egress

被引：111

作者：

Mendonca, Luiza ^{[1
]}

Howe, Andrew ^{[2
]}

Gilchrist, James B. ^{[2
]}

Sheng, Yuewen ^{[2
]}

Sun, Dapeng ^{[3
]}

Knight, Michael L. ^{[4
]}

Zanetti-Domingues, Laura C. ^{[5
]}

Bateman, Benji ^{[5
]}

Krebs, Anna-Sophia ^{[1
]}

Chen, Long ^{[1
]}

Radecke, Julika ^{[2
]}

Li, Vivian D. ^{[6
]}

Ni, Tao ^{[1
]}

Kounatidis, Ilias ^{[2
]}

Koronfel, Mohamed A. ^{[2
]}

Szynkiewicz, Marta ^{[5
]}

Harkiolaki, Maria ^{[2
]}

Martin-Fernandez, Marisa L. ^{[5
]}

James, William ^{[4
]}

Zhang, Peijun ^{[1
,2
,3
]}

机构：

[1] Univ Oxford, Div Struct Biol, Wellcome Trust Ctr Human Genet, Oxford, England

[2] Harwell Sci & Innovat Campus, Diamond Light Source, Didcot, Oxon, England

[3] Univ Pittsburgh, Dept Biol Struct, Pittsburgh, PA 15260 USA

[4] Univ Oxford, Sir William Dunn Sch Pathol, Oxford, England

[5] Rutherford Appleton Lab, Sci & Technol Facil Council, Cent Laser Facil, Didcot, Oxon, England

[6] Univ Cambridge, Murray Edwards Coll, Cambridge, England

来源：

NATURE COMMUNICATIONS | 2021年 / 12卷 / 01期

基金：

美国国家卫生研究院; 英国生物技术与生命科学研究理事会; 英国惠康基金;

关键词：

CORONAVIRUS REPLICATION; VISUALIZATION; ACTIVATION; PROTEINS; ENTRY; RNA;

D O I：

10.1038/s41467-021-24887-y

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

In this study, Peijun Zhang and colleagues use cryoFIB/SEM volume imaging and soft x-ray cryo-tomography with cryo-electron tomography (cryoET) of cellular periphery, lamellae, and subtomogram averaging to place critical structural events in the SARS-CoV-2 infection cycle in the context of whole-cell images. Since the outbreak of the SARS-CoV-2 pandemic, there have been intense structural studies on purified viral components and inactivated viruses. However, structural and ultrastructural evidence on how the SARS-CoV-2 infection progresses in the native cellular context is scarce, and there is a lack of comprehensive knowledge on the SARS-CoV-2 replicative cycle. To correlate cytopathic events induced by SARS-CoV-2 with virus replication processes in frozen-hydrated cells, we established a unique multi-modal, multi-scale cryo-correlative platform to image SARS-CoV-2 infection in Vero cells. This platform combines serial cryoFIB/SEM volume imaging and soft X-ray cryo-tomography with cell lamellae-based cryo-electron tomography (cryoET) and subtomogram averaging. Here we report critical SARS-CoV-2 structural events - e.g. viral RNA transport portals, virus assembly intermediates, virus egress pathway, and native virus spike structures, in the context of whole-cell volumes revealing drastic cytppathic changes. This integrated approach allows a holistic view of SARS-CoV-2 infection, from the whole cell to individual molecules.

引用

页数：10

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