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A molecular pore spans the double membrane of the coronavirus replication organelle

View ORCID ProfileGeorg Wolff, View ORCID ProfileRonald W.A.L. Limpens, Jessika C. Zevenhoven-Dobbe, Ulrike Laugks, Shawn Zheng, View ORCID ProfileAnja W. M. de Jong, View ORCID ProfileRoman I. Koning, View ORCID ProfileDavid A. Agard, View ORCID ProfileKay Grünewald, View ORCID ProfileAbraham J. Koster, View ORCID ProfileEric J. Snijder, View ORCID ProfileMontserrat Bárcena
doi: https://doi.org/10.1101/2020.06.25.171686
Georg Wolff
1Department of Cell and Chemical Biology, Section Electron Microscopy, Leiden University Medical Center, Leiden 2333 ZC, The Netherlands
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Ronald W.A.L. Limpens
1Department of Cell and Chemical Biology, Section Electron Microscopy, Leiden University Medical Center, Leiden 2333 ZC, The Netherlands
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Jessika C. Zevenhoven-Dobbe
2Department of Medical Microbiology, Molecular Virology Laboratory, Leiden University Medical Center, Leiden 2333 ZA, The Netherlands
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Ulrike Laugks
3Department of Structural Cell Biology of Viruses, Centre for Structural Systems Biology, Heinrich Pette Institute, Leibnitz Institute of Experimental Virology, University of Hamburg, Hamburg, Germany
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Shawn Zheng
4Department of Biochemistry and Biophysics, Howard Hughes Medical Institute, University of California San Francisco, San Francisco, CA 94143, United States
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Anja W. M. de Jong
1Department of Cell and Chemical Biology, Section Electron Microscopy, Leiden University Medical Center, Leiden 2333 ZC, The Netherlands
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Roman I. Koning
1Department of Cell and Chemical Biology, Section Electron Microscopy, Leiden University Medical Center, Leiden 2333 ZC, The Netherlands
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David A. Agard
5Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, CA 94143, United States
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Kay Grünewald
3Department of Structural Cell Biology of Viruses, Centre for Structural Systems Biology, Heinrich Pette Institute, Leibnitz Institute of Experimental Virology, University of Hamburg, Hamburg, Germany
6Department of Chemistry, MIN Faculty, Universität Hamburg, Hamburg, Germany
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Abraham J. Koster
1Department of Cell and Chemical Biology, Section Electron Microscopy, Leiden University Medical Center, Leiden 2333 ZC, The Netherlands
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Eric J. Snijder
2Department of Medical Microbiology, Molecular Virology Laboratory, Leiden University Medical Center, Leiden 2333 ZA, The Netherlands
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Montserrat Bárcena
1Department of Cell and Chemical Biology, Section Electron Microscopy, Leiden University Medical Center, Leiden 2333 ZC, The Netherlands
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  • For correspondence: m.barcena@lumc.nl
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Abstract

Coronavirus genome replication is associated with virus-induced cytosolic double-membrane vesicles, which may provide a tailored micro-environment for viral RNA synthesis in the infected cell. However, it is unclear how newly synthesized genomes and mRNAs can travel from these sealed replication compartments to the cytosol to ensure their translation and the assembly of progeny virions. Here, using cellular electron cryo-microscopy, we unveiled a molecular pore complex that spans both membranes of the double-membrane vesicle and would allow export of RNA to the cytosol. A hexameric assembly of a large viral transmembrane protein was found to form the core of the crown-shaped complex. This coronavirus-specific structure likely plays a critical role in coronavirus replication and thus constitutes a novel drug target

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted June 25, 2020.
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A molecular pore spans the double membrane of the coronavirus replication organelle
Georg Wolff, Ronald W.A.L. Limpens, Jessika C. Zevenhoven-Dobbe, Ulrike Laugks, Shawn Zheng, Anja W. M. de Jong, Roman I. Koning, David A. Agard, Kay Grünewald, Abraham J. Koster, Eric J. Snijder, Montserrat Bárcena
bioRxiv 2020.06.25.171686; doi: https://doi.org/10.1101/2020.06.25.171686
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A molecular pore spans the double membrane of the coronavirus replication organelle
Georg Wolff, Ronald W.A.L. Limpens, Jessika C. Zevenhoven-Dobbe, Ulrike Laugks, Shawn Zheng, Anja W. M. de Jong, Roman I. Koning, David A. Agard, Kay Grünewald, Abraham J. Koster, Eric J. Snijder, Montserrat Bárcena
bioRxiv 2020.06.25.171686; doi: https://doi.org/10.1101/2020.06.25.171686

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