TY - JOUR
T1 - Correlative multi-scale cryo-imaging unveils SARS-CoV-2 assembly and egress
AU - Mendonça, Luiza
AU - Howe, Andrew
AU - Gilchrist, James B.
AU - Sheng, Yuewen
AU - Sun, Dapeng
AU - Knight, Michael L.
AU - Zanetti-Domingues, Laura C.
AU - Bateman, Benji
AU - Krebs, Anna Sophia
AU - Chen, Long
AU - Radecke, Julika
AU - Li, Vivian D.
AU - Ni, Tao
AU - Kounatidis, Ilias
AU - Koronfel, Mohamed A.
AU - Szynkiewicz, Marta
AU - Harkiolaki, Maria
AU - Martin-Fernandez, Marisa L.
AU - James, William
AU - Zhang, Peijun
N1 - Publisher Copyright:
© 2021, The Author(s).
PY - 2021/12/1
Y1 - 2021/12/1
N2 - 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.
AB - 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.
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U2 - 10.1038/s41467-021-24887-y
DO - 10.1038/s41467-021-24887-y
M3 - Article
C2 - 34330917
AN - SCOPUS:85111625783
SN - 2041-1723
VL - 12
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 4629
ER -
