Journal
JOURNAL OF BIOLOGICAL CHEMISTRY
Volume 295, Issue 15, Pages 4773-4779Publisher
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
DOI: 10.1074/jbc.AC120.013056
Keywords
plus-stranded RNA virus; viral polymerase; drug development; enzyme inhibitor; nucleoside; nucleotide analog; coronavirus; positive-sense RNA virus; Ebola virus (EBOV); Middle East respiratory syndrome coronavirus (MERS?CoV); SARS?CoV-2; remdesivir; antiviral drug; RNA chain termination; RNA-dependent RNA polymerase (RdRp); viral replicase
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Funding
- Canadian Institutes of Health Research [159507]
- Alberta Ministry of Economic Development, Trade and Tourism by the Major Innovation Fund Program for the AMR-One Health Consortium
- Gilead Sciences
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Antiviral drugs for managing infections with human coronaviruses are not yet approved, posing a serious challenge to current global efforts aimed at containing the outbreak of severe acute respiratory syndrome?coronavirus 2 (CoV-2). Remdesivir (RDV) is an investigational compound with a broad spectrum of antiviral activities against RNA viruses, including severe acute respiratory syndrome?CoV and Middle East respiratory syndrome (MERS?CoV). RDV is a nucleotide analog inhibitor of RNA-dependent RNA polymerases (RdRps). Here, we co-expressed the MERS?CoV nonstructural proteins nsp5, nsp7, nsp8, and nsp12 (RdRp) in insect cells as a part a polyprotein to study the mechanism of inhibition of MERS?CoV RdRp by RDV. We initially demonstrated that nsp8 and nsp12 form an active complex. The triphosphate form of the inhibitor (RDV-TP) competes with its natural counterpart ATP. Of note, the selectivity value for RDV-TP obtained here with a steady-state approach suggests that it is more efficiently incorporated than ATP and two other nucleotide analogs. Once incorporated at position i, the inhibitor caused RNA synthesis arrest at position i + 3. Hence, the likely mechanism of action is delayed RNA chain termination. The additional three nucleotides may protect the inhibitor from excision by the viral 3??5? exonuclease activity. Together, these results help to explain the high potency of RDV against RNA viruses in cell-based assays.
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