In situ structure of virus capsids within cell nuclei by correlative light and cryo-electron tomography

Vijayakrishnan, S. , McElwee, M., Loney, C. , Rixon, F. and Bhella, D. (2020) In situ structure of virus capsids within cell nuclei by correlative light and cryo-electron tomography. Scientific Reports, 10, 17596. (doi: 10.1038/s41598-020-74104-x) (PMID:33077791) (PMCID:PMC7572381)

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Cryo electron microscopy (cryo-EM), a key method for structure determination involves imaging purified material embedded in vitreous ice. Images are then computationally processed to obtain three-dimensional structures approaching atomic resolution. There is increasing interest in extending structural studies by cryo-EM into the cell, where biological structures and processes may be imaged in context. The limited penetrating power of electrons prevents imaging of thick specimens (> 500 nm) however. Cryo-sectioning methods employed to overcome this are technically challenging, subject to artefacts or involve specialised and costly equipment. Here we describe the first structure of herpesvirus capsids determined by sub-tomogram averaging from nuclei of eukaryotic cells, achieved by cryo-electron tomography (cryo-ET) of re-vitrified cell sections prepared using the Tokuyasu method. Our reconstructions confirm that the capsid associated tegument complex is present on capsids prior to nuclear egress. We demonstrate that this method is suited to both 3D structure determination and correlative light/electron microscopy, thus expanding the scope of cryogenic cellular imaging.

Item Type:Articles
Additional Information:This work was supported by the Medical Research Council (Grant number: MC_UU_12014/7).
Glasgow Author(s) Enlighten ID:Vijayakrishnan, Dr Swetha and Bhella, Professor David and Loney, Mr Colin and McElwee, Dr Marion and Rixon, Dr Frazer
Creator Roles:
Vijayakrishnan, S.Conceptualization, Project administration, Investigation, Methodology, Formal analysis, Visualization, Writing – original draft, Writing – review and editing
McElwee, M.Investigation, Resources
Loney, C.Investigation, Resources, Methodology
Rixon, F.Conceptualization, Investigation, Methodology, Resources
Bhella, D.Conceptualization, Investigation, Methodology, Formal analysis, Funding acquisition, Writing – review and editing
Authors: Vijayakrishnan, S., McElwee, M., Loney, C., Rixon, F., and Bhella, D.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Scientific Reports
Publisher:Nature Research
ISSN (Online):2045-2322
Copyright Holders:Copyright © 2020 The Authors
First Published:First published in Scientific Reports 10:17596
Publisher Policy:Reproduced under a Creative Commons License

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