SKM 2021 – wissenschaftliches Programm

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BP: Fachverband Biologische Physik

BP 3: Protein Structure and Dynamics

BP 3.1: Hauptvortrag

Donnerstag, 30. September 2021, 10:00–10:30, H1

SARS-CoV-2 induced membrane remodeling in infected cells revealed by in celullo cryo-ET — Steffen Klein, Liv Zimmermann, Sophie Winter, Mirko Cortese, Moritz Wachsmuth-Melm, Christopher Neufeldt, Berati Cerikan, Megan Stanifer, Steeve Boulant, Ralf Bartenschlager, and •Petr Chlanda — Heidelberg University Hospital, Heidelberg, Germany

Coronavirus replication in the host cell causes extensive remodeling of cellular membranes. To better understand the governing mechanisms of SARS-CoV-2 membrane remodeling during RNA replication and virus assembly we visualized hubs of virus replication and assembly using cryo-electron tomography of infected cells. Our data reveal the architecture of double-membrane vesicles which are associated with viral genome replication. Viral RNA filaments inside these compartments show a diameter consistent with double-stranded RNA and displayed frequent branching, likely representing secondary structures. Virion assembly sites were found at cisternae enriched in spike trimers and viral ribonucleoprotein complexes (vRNPs) at the cytoplasmic side. We further structurally analyzed the viral genome in newly assembled virions and revealed that the viral RNA is encased by multiple individual cylindrical vRNPs. We propose that this arrangement allows the incorporation of the unusually large coronavirus genome into the virion while maintaining high steric flexibility between the vRNPs during virion assembly.