In situ structure of virus capsids within cell nuclei by correlative light and cryo-electron tomography.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
19 10 2020
19 10 2020
Historique:
received:
28
02
2020
accepted:
24
09
2020
entrez:
20
10
2020
pubmed:
21
10
2020
medline:
15
1
2021
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 33077791
doi: 10.1038/s41598-020-74104-x
pii: 10.1038/s41598-020-74104-x
pmc: PMC7572381
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
17596Subventions
Organisme : Medical Research Council
ID : MC_UU_12014/6
Pays : United Kingdom
Organisme : Medical Research Council
ID : MC_UU_12014/7
Pays : United Kingdom
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