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PreprintCited by 464SARS-CoV-2 and COVID-19 ResearchAnimal Virus Infections StudiesVirology and Viral Diseases

A unifying structural and functional model of the coronavirus replication organelle: tracking down RNA synthesis

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Snijder, Eric J., Limpens, Ronald W.A.L., de Wilde, Adriaan H., de Jong, Anja W. M., Zevenhoven-Dobbe, Jessika C., Maier, Helena J., Faas, Frank F.G.A., Koster, Abraham J. +1 more·2020·PLoS Biology
DOI 10.1101/2020.03.24.005298PMC PMC7302735

Preprint — published as 10.1371/journal.pbio.3000715 · Paper page

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Abstract

Zoonotic coronavirus (CoV) infections, like those responsible for the current SARS-CoV-2 epidemic, cause grave international public health concern. In infected cells, the CoV RNA-synthesizing machinery associates with modified endoplasmic reticulum membranes that are transformed into the viral replication organelle (RO). While double-membrane vesicles (DMVs) appear to be a pan -coronavirus RO element, studies to date describe an assortment of additional coronavirus-induced membrane structures. Despite much speculation, it remains unclear which RO element(s) accommodate viral RNA synthesis. Here we provide detailed 2D and 3D analyses of CoV ROs and show that diverse CoVs essentially induce the same membrane modifications, including the small open double-membrane spherules (DMSs) previously thought to be restricted to gamma- and delta-CoV infections and proposed as sites of replication. Metabolic labelling of newly-synthesized viral RNA followed by quantitative EM autoradiography revealed abundant viral RNA synthesis associated with DMVs in cells infected with the beta-CoVs MERS-CoV and SARS-CoV, and the gamma-CoV infectious bronchitis virus. RNA synthesis could not be linked to DMSs or any other cellular or virus-induced structure. Our results provide a unifying model of the CoV RO and clearly establish DMVs as the central hub for viral RNA synthesis and a potential drug target in coronavirus infection.

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Authors
Snijder, E. J., Limpens, R. W. A. L., de Wilde, A. H., de Jong, A. W. M., Zevenhoven-Dobbe, J. C., Maier, H. J., Faas, F. F. G. A., Koster, A. J., & Bárcena, M.
Affiliations
  • Leiden University Medical Center (NL)
  • The Pirbright Institute (GB)
Corresponding author
  • Snijder, Eric J.
  • Bárcena, Montserrat
ORCID
Venue
PLoS Biology
Publisher
openRxiv
Published
2020-03-24
Volume
18
Issue
6
Pages
e3000715-e3000715
ISSN
1544-9173 (ISSN-L)
DOI
10.1101/2020.03.24.005298
PMC
PMC7302735
OpenAlex
W3033917173
Cited by
464
References
0
Citations by year
2026: 44 · 2025: 36 · 2024: 52 · 2023: 74 · 2022: 89 · 2021: 116 · 2020: 51 · 2019: 2
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10.48
Type
journal article
Language
en
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  • PLoS Biology (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
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  • Leiden Repository (Leiden University) (submittedVersion, cc-by)
  • Data Archiving and Networked Services (DANS) (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Informationszentrum Lebenswissenschaften (ZB MED) (submittedVersion, cc-by)
Funders
  • Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO-MEERVOUD-836.10.003, NWO-MEERVOUD-863.10.003, 700-57-301, MEERVOUD-836.10.003, 836.10.003, 863.10.003)
  • Biotechnology and Biological Sciences Research Council (BBS/E/I/00007034, BB/N002350/1, BBS/E/I/COV07001)
Topics
  • Health Sciences › Medicine › Infectious Diseases › SARS-CoV-2 and COVID-19 Research
  • Life Sciences › Agricultural and Biological Sciences › Animal Science and Zoology › Animal Virus Infections Studies
  • Health Sciences › Medicine › Epidemiology › Virology and Viral Diseases
Keywords
viral RNA synthesis, double-membrane vesicles, SARS-CoV-2, MERS-CoV, SARS-CoV, infectious bronchitis virus, betacoronaviruses, antiviral drug targets
MeSH terms
Betacoronavirus, COVID-19, SARS-CoV-2, Animals, Cell Line, Chlorocebus aethiops, Endoplasmic Reticulum, Humans, Pneumonia, Viral, RNA, Viral, Vero Cells, Virus Replication, Coronavirus, Coronavirus Infections, Electron Microscope Tomography, Pandemics, Middle East Respiratory Syndrome Coronavirus

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

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