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)

Abstract

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).
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Rixon, Dr Frazer and Bhella, Professor David and Loney, Mr Colin and McElwee, Dr Marion and Vijayakrishnan, Dr Swetha

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 > School of Infection & Immunity; College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Virus Research
• Journal Name: Scientific Reports
• Publisher: Nature Research
• ISSN: 2045-2322
• 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

Project Code	Award No	Project Name	Principal Investigator	Funder's Name	Funder Ref	Lead Dept
					MC_UU_12014/7