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

Zoonotic coronavirus (CoV) infections, such as those responsible for the current severe acute respiratory syndrome-CoV 2 (SARS-CoV-2) pandemic, cause grave international public health concern. In infected cells, the CoV RNA-synthesizing machinery associates with modified endoplasmic reticulum membra...

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Autores principales: 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., Bárcena, Montserrat
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7302735/
https://www.ncbi.nlm.nih.gov/pubmed/32511245
http://dx.doi.org/10.1371/journal.pbio.3000715
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author 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.
Bárcena, Montserrat
author_facet 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.
Bárcena, Montserrat
author_sort Snijder, Eric J.
collection PubMed
description Zoonotic coronavirus (CoV) infections, such as those responsible for the current severe acute respiratory syndrome-CoV 2 (SARS-CoV-2) pandemic, 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). Although double-membrane vesicles (DMVs) appear to be a pan-CoV RO element, studies to date describe an assortment of additional CoV-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 labeling of newly synthesized viral RNA followed by quantitative electron microscopy (EM) autoradiography revealed abundant viral RNA synthesis associated with DMVs in cells infected with the beta-CoVs Middle East respiratory syndrome-CoV (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 CoV infection.
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spelling pubmed-73027352020-06-19 A unifying structural and functional model of the coronavirus replication organelle: Tracking down RNA synthesis 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. Bárcena, Montserrat PLoS Biol Research Article Zoonotic coronavirus (CoV) infections, such as those responsible for the current severe acute respiratory syndrome-CoV 2 (SARS-CoV-2) pandemic, 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). Although double-membrane vesicles (DMVs) appear to be a pan-CoV RO element, studies to date describe an assortment of additional CoV-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 labeling of newly synthesized viral RNA followed by quantitative electron microscopy (EM) autoradiography revealed abundant viral RNA synthesis associated with DMVs in cells infected with the beta-CoVs Middle East respiratory syndrome-CoV (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 CoV infection. Public Library of Science 2020-06-08 /pmc/articles/PMC7302735/ /pubmed/32511245 http://dx.doi.org/10.1371/journal.pbio.3000715 Text en © 2020 Snijder et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
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.
Bárcena, Montserrat
A unifying structural and functional model of the coronavirus replication organelle: Tracking down RNA synthesis
title A unifying structural and functional model of the coronavirus replication organelle: Tracking down RNA synthesis
title_full A unifying structural and functional model of the coronavirus replication organelle: Tracking down RNA synthesis
title_fullStr A unifying structural and functional model of the coronavirus replication organelle: Tracking down RNA synthesis
title_full_unstemmed A unifying structural and functional model of the coronavirus replication organelle: Tracking down RNA synthesis
title_short A unifying structural and functional model of the coronavirus replication organelle: Tracking down RNA synthesis
title_sort unifying structural and functional model of the coronavirus replication organelle: tracking down rna synthesis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7302735/
https://www.ncbi.nlm.nih.gov/pubmed/32511245
http://dx.doi.org/10.1371/journal.pbio.3000715
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