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

DOI: 10.1038/s41467-021-24887-y

Authors: Luiza Mendonca (Wellcome Trust Centre for Human Genetics, University of Oxford) , Andrew Howe (Diamond Light Source) , James B. Gilchrist (Diamond Light Source) , Yuewen Sheng (Diamond Light Source) , Dapeng Sun (University of Pittsburgh) , Michael L. Knight (University of Oxford) , Laura C. Zanetti-Domingues (Central Laser Facility, Science and Technology Facilities Council) , Benji Bateman (Central Laser Facility, Science and Technology Facilities Council) , Anna-Sophia Krebs (Wellcome Trust Centre for Human Genetics, University of Oxford) , Long Chen (Wellcome Trust Centre for Human Genetics, University of Oxford) , Julika Radecke (Diamond Light Source) , Vivian D. Li (University of Cambridge) , Tao Ni (Wellcome Trust Centre for Human Genetics, University of Oxford) , Ilias Kounatidis (Diamond Light Source) , Mohamed A. Koronfel (Diamond Light Source) , Marta Szynkiewicz (Central Laser Facility, Science and Technology Facilities Council) , Maria Harkiolaki (Diamond Light Source) , Marisa Martin-Fernandez (Central Laser Facility, Science and Technology Facilities Council) , William James (University of Oxford) , Peijun Zhang (Wellcome Trust Centre for Human Genetics, University of Oxford; Diamond Light Source; University of Pittsburgh)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature Communications , VOL 12

State: Published (Approved)
Published: July 2021
Diamond Proposal Number(s): 21004 , 26987

Abstract: 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.

Journal Keywords: Cryoelectron tomography; Pathogens; SARS-CoV-2; Virus structures

Diamond Keywords: COVID-19; Viruses

Subject Areas: Biology and Bio-materials

Diamond Offline Facilities: Electron Bio-Imaging Centre (eBIC)
Instruments: B24-Cryo Soft X-ray Tomography , Krios II-Titan Krios II at Diamond

Added On: 02/08/2021 14:06

Documents:
s41467-021-24887-y.pdf

Discipline Tags:

Pathogens Infectious Diseases Health & Wellbeing Structural biology Life Sciences & Biotech

Technical Tags:

Imaging Microscopy Tomography Electron Microscopy (EM) Cryo Electron Microscopy (Cryo EM) Cryo-soft X-ray Tomography (Cryo-SXT)