tetano
Editor, Senior Moderator
J Biol Chem. 2020 Feb 24. pii: jbc.AC120.013056. doi: 10.1074/jbc.AC120.013056. [Epub ahead of print] [h=1]The antiviral compound remdesivir potently inhibits RNA-dependent RNA polymerase from Middle East respiratory syndrome coronavirus.[/h]
Gordon CJ[SUP]1[/SUP], Tchesnokov EP[SUP]1[/SUP], Feng JY[SUP]2[/SUP], Porter DP[SUP]3[/SUP], Gotte M[SUP]4[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] 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 (SARS-CoV-2). Remdesivir (RDV) is an investigational compound with a broad spectrum of antiviral activities against RNA viruses, including SARS-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 analogues. 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.
Published under license by The American Society for Biochemistry and Molecular Biology, Inc.
[h=4]KEYWORDS:[/h] Ebola virus (EBOV); Middle East respiratory syndrome coronavirus (MERS-CoV), SARS-CoV-2; RNA-dependent RNA polymerase (RdRp), viral replicase; coronavirus, positive-sense RNA virus; drug development; enzyme inhibitor; nucleoside/nucleotide analogue; plus-stranded RNA virus; remdesivir, antiviral drug, RNA chain-termination; viral polymerase
PMID: 32094225 DOI: 10.1074/jbc.AC120.013056
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Gordon CJ[SUP]1[/SUP], Tchesnokov EP[SUP]1[/SUP], Feng JY[SUP]2[/SUP], Porter DP[SUP]3[/SUP], Gotte M[SUP]4[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] 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 (SARS-CoV-2). Remdesivir (RDV) is an investigational compound with a broad spectrum of antiviral activities against RNA viruses, including SARS-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 analogues. 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.
Published under license by The American Society for Biochemistry and Molecular Biology, Inc.
[h=4]KEYWORDS:[/h] Ebola virus (EBOV); Middle East respiratory syndrome coronavirus (MERS-CoV), SARS-CoV-2; RNA-dependent RNA polymerase (RdRp), viral replicase; coronavirus, positive-sense RNA virus; drug development; enzyme inhibitor; nucleoside/nucleotide analogue; plus-stranded RNA virus; remdesivir, antiviral drug, RNA chain-termination; viral polymerase
PMID: 32094225 DOI: 10.1074/jbc.AC120.013056
Free full text