Interferon-gamma induced RNF213 targets nascent double membrane vesicles to inhibit coronavirus primary translation

Innate immune systems employ many mechanisms to inhibit the replication of viruses that profoundly affect host range and pathogenicity. To identify proteins that restrict human respiratory virus replication, we executed CRISPR screens targeting both type-I and type-II interferon stimulated genes. Among the candidate antiviral genes identified, the giant ring finger protein 213 (RNF213) was a key mediator of antiviral activity of interferon gamma (IFN{gamma}) against the human betacoronavirus HCoV-OC43. RNF213 has ATPase and ubiquitin ligase activities, both of which were required for anti-HCoV-OC43 activity. Using newly designed reporter viruses, we demonstrate that RNF213 acts by a previously undescribed mechanism, inhibiting the translation of incoming viral genomes. RNF213 colocalized and coimmunoprecipitated with the primary viral translation product, NSP3, that orchestrates the formation coronavirus replication compartments. Notably, RNF213 restricted genetically diverse common human coronaviruses, but SARS-CoV-2 apparently evades interaction with and inhibition by RNF213.