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SARS-CoV-2 structure and replication characterized by in situ cryo-electron tomography

Open access
Klein, Steffen, Cortese, Mirko, Winter, Sophie L., Wachsmuth-Melm, Moritz, Neufeldt, Christopher J., Cerikan, Berati, Stanifer, Megan L., Boulant, Steeve, Bartenschlager, Ralf, Chlanda, Petr·2020·Nature Communications

Published version of a preprint

Abstract

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of the COVID19 pandemic, is a highly pathogenic β-coronavirus. As other coronaviruses, SARS-CoV-2 is enveloped, replicates in the cytoplasm and assembles at intracellular membranes. Here, we structurally characterize the viral replication compartment and report critical insights into the budding mechanism of the virus, and the structure of extracellular virions close to their native state by in situ cryo-electron tomography and subtomogram averaging. We directly visualize RNA filaments inside the double membrane vesicles, compartments associated with viral replication. The RNA filaments show a diameter consistent with double-stranded RNA and frequent branching likely representing RNA secondary structures. We report that assembled S trimers in lumenal cisternae do not alone induce membrane bending but laterally reorganize on the envelope during virion assembly. The viral ribonucleoprotein complexes (vRNPs) are accumulated at the curved membrane characteristic for budding sites suggesting that vRNP recruitment is enhanced by membrane curvature. Subtomogram averaging shows that vRNPs are distinct cylindrical assemblies. We propose that the genome is packaged around multiple separate vRNP complexes, thereby allowing incorporation of the unusually large coronavirus genome into the virion while maintaining high steric flexibility between the vRNPs.

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Research works printed but not linked: 3

  1. Escors, D., Ortego, J., Laude, H. & Enjuanes, L. The membrane M protein carboxy terminus binds to transmissible gastroenteritis coronavirus core and contributes to core stability. J. Virol. 75, 1312–1324 (2001). · doi:10.1128/jvi.75.3.1312-1324.2001
  2. McIntosh, K. & Perlman, S. in Mandell, Douglas, and Bennett’s Principles and Practice of Infectious Diseases, Updated Edition 8th edn (Elsevier Saunders, Philadelphia, 2015).
  3. Beltrán, C., Cucker, F., Krick, T., Pinkus, A. & Szanto, A. in Foundations of Computational Mathematics, Budapest 2011 Ch. 1, 1–15 (Cambridge Univ. Press, 2012). · doi:10.1017/cbo9781139095402

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Metadata

Authors
Klein, S., Cortese, M., Winter, S. L., Wachsmuth-Melm, M., Neufeldt, C. J., Cerikan, B., Stanifer, M. L., Boulant, S., Bartenschlager, R., & Chlanda, P.
Venue
Nature Communications
Published
2020-11-18
Volume
11
Issue
1
Pages
5885-5885
ISSN
2041-1723 (ISSN-L)
OpenAlex
W3103021984
Cited by
731
References
56
FWCI
15.03
Type
journal article
Language
en
Open access
gold
Licence
Funders
  • CHS Stiftung
  • Deutsche Forschungsgemeinschaft (SFB1129-240245660)

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CoChat. (2026). Citation check: SARS-CoV-2 structure and replication characterized by in situ cryo-electron tomography. https://app.cochat.ai/papers/10.1038/s41467-020-19619-7
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Klein, S., Cortese, M., Winter, S. L., Wachsmuth-Melm, M., Neufeldt, C. J., Cerikan, B., Stanifer, M. L., Boulant, S., Bartenschlager, R., & Chlanda, P. (2020). SARS-CoV-2 structure and replication characterized by in situ cryo-electron tomography. Nature Communications. https://doi.org/10.1038/s41467-020-19619-7

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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).

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