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Abstract

Coronavirus Replication in Host Cells

A Molecular Pore Spans the Double Membrane of the Replication Organelle

Within the last 2 decades, there have been 3 potentially lethal coronavirus outbreaks for humans including SARS-CoV-2 which causes COVID-19. 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.

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For this study, cellular electron cryo-microscopy was used to visualize a molecular pore complex that spans both membranes of the 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 structure likely plays a critical role in replication and constitutes a potential drug target. SARS-CoV-2 infected VeroE6 cells used for the study were prepared for electron cryo-tomography in part with an automated plunge freezer from Leica Microsystems. Refer to the publication for more details.

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

G. Wolff, R.W.A.L. Limpens, J.C. Zevenhoven-Dobbe, U. Laugks, S. Zheng, A.W.M. de Jong, R.I. Koning, D.A. Agard, K. Grünewald, A.J. Koster, E.J. Snijder, M. Bárcena

Science (2020) eabd3629, DOI: 10.1126/science.abd3629