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A Coronaviral Pore-Replicase Complex Drives RNA Synthesis in Double Membrane Vesicles

bioRxiv, ISSN: 2692-8205
2024
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Article Description

Coronavirus-infected cells contain double-membrane vesicles (DMVs) that are key for viral RNA replication and transcription, perforated by hexameric pores connecting the vesicular lumen to the cytoplasm. How pores form and traverse two membranes, and how DMVs organize RNA synthesis, is unknown. Using structure prediction and functional assays, we show that the non-structural viral membrane protein nsp4 is the key DMV pore organizer, spanning the double membrane and forming most of the pore lining. Nsp4 interacts with nsp3 on the cytoplasmic side and with the viral replicase inside the DMV. Newly synthesized mRNAs exit the DMV into the cytoplasm, passing through a narrow ring of conserved nsp4 residues. Steric constraints imposed by the ring predict that modified nucleobases block mRNA transit, with broad spectrum anti-coronaviral activity.

Bibliographic Details

Anan Chen; Ana Mihaela Lupan; Taciani de Almeida Magalhães; Adrian Salic; Rui Tong Quek; Kierra S. Hardy; Pamela A. Silver; Timothy J. Mitchison; Stefan G. Stanciu; Mihaela Asaftei; George A. Stanciu

Cold Spring Harbor Laboratory

Biochemistry, Genetics and Molecular Biology; Agricultural and Biological Sciences; Immunology and Microbiology; Neuroscience; Pharmacology, Toxicology and Pharmaceutics

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