dc.contributor.author | Gordon, Calvin J. et al. | |
dc.date.accessioned | 2020-04-23T18:48:00Z | |
dc.date.available | 2020-04-23T18:48:00Z | |
dc.date.issued | 2020-04-13 | |
dc.identifier.uri | https://doi.org/10.1074/jbc.ra120.013679 | en_US |
dc.identifier.uri | https://hdl.handle.net/20.500.12663/1185 | |
dc.description.abstract | Effective treatments for coronavirus disease 2019 (COVID-19) are urgently needed to control this current pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Replication of SARS-CoV-2 depends on the viral RNA-dependent RNA polymerase (RdRp), which is the likely target of the investigational nucleotide analogue remdesivir (RDV). RDV shows broad-spectrum antiviral activity against RNA viruses, and previous studies with RdRps from Ebola virus (EBOV) and Middle East respiratory syndrome coronavirus (MERS-CoV) have revealed that delayed chain-termination is RDV’s plausible mechanism of action. Here, we expressed and purified active SARS-CoV-2 RdRp composed of the non-structural proteins nsp8 and nsp12. Enzyme kinetics indicated that this RdRp efficiently incorporates the active triphosphate form of RDV (RDV-TP) into RNA. Incorporation of RDV-TP at position i caused termination of RNA synthesis at position i+3. We obtained almost identical results with SARS-CoV, MERS-CoV, and SARS-CoV-2 RdRps. A unique property of RDV-TP is its high selectivity over incorporation of its natural nucleotide counterpart ATP. In this regard, the triphosphate forms of 2’-C–methylated compounds, including sofosbuvir, approved for the management of hepatitis C virus infection, and the broad-acting antivirals favipiravir and ribavirin, exhibited significant deficits. Furthermore, we provide evidence for the target specificity of RDV, as RDV-TP was less efficiently incorporated by the distantly related Lassa virus RdRp, and termination of RNA synthesis was not observed. These results collectively provide a unifying, refined mechanism of RDV-mediated RNA synthesis inhibition in coronaviruses and define this nucleotide analogue as a direct-acting antiviral (DAA). | en_US |
dc.language | English | en_US |
dc.subject | COVID-19 | en_US |
dc.subject | Coronavirus | en_US |
dc.subject | Infectious Diseases | en_US |
dc.subject | Middle East Respiratory Syndrome Coronavirus | en_US |
dc.subject | Severe Acute Respiratory Syndrome | en_US |
dc.subject | SARS-CoV-2 | en_US |
dc.subject | Ebolavirus | en_US |
dc.subject | Lassa virus | en_US |
dc.subject | DNA-Directed RNA Polymerases | en_US |
dc.subject | Sofosbuvir | en_US |
dc.subject | Remdesivir | en_US |
dc.title | Remdesivir is a direct-acting antiviral that inhibits RNA-dependent RNA polymerase from severe acute respiratory syndrome coronavirus 2 with high potency | en_US |
eihealth.country | Others | en_US |
eihealth.category | Candidate therapeutics RD | en_US |
eihealth.type | Other publications | en_US |
eihealth.maincategory | Save Lives / Salvar Vidas | en_US |
dc.relation.ispartofjournal | Journal of Biological Chemistry | en_US |