The antiviral compound remdesivir potently inhibits RNA-dependent RNA polymerase from Middle East respiratory syndrome coronavirus
Identifieur interne : 000051 ( Main/Exploration ); précédent : 000050; suivant : 000052The antiviral compound remdesivir potently inhibits RNA-dependent RNA polymerase from Middle East respiratory syndrome coronavirus
Auteurs : Calvin J. Gordon [Canada] ; Egor P. Tchesnokov [Canada] ; Joy Y. Feng [États-Unis] ; Danielle P. Porter [États-Unis] ; Matthias Götte [Canada]Source :
- The Journal of Biological Chemistry [ 0021-9258 ] ; 2020.
Descripteurs français
- KwdFr :
- AMP (), AMP (analogues et dérivés), AMP (pharmacologie), ARN, Alanine (), Alanine (analogues et dérivés), Alanine (pharmacologie), Animaux, Antiviraux (), Antiviraux (pharmacologie), Cellules Sf9, Coronavirus (enzymologie), Coronavirus du syndrome respiratoire du Moyen-Orient (enzymologie), Ebolavirus (enzymologie), Expression des gènes, Inhibiteurs de la synthèse d'acide nucléique (), Inhibiteurs de la synthèse d'acide nucléique (pharmacologie), Protéines virales non structurales (génétique), RNA replicase (antagonistes et inhibiteurs), RNA replicase (génétique), Réplication virale ().
- MESH :
- analogues et dérivés : AMP, Alanine.
- antagonistes et inhibiteurs : RNA replicase.
- enzymologie : Coronavirus, Coronavirus du syndrome respiratoire du Moyen-Orient, Ebolavirus.
- génétique : Protéines virales non structurales, RNA replicase.
- pharmacologie : AMP, Alanine, Antiviraux, Inhibiteurs de la synthèse d'acide nucléique.
- AMP, ARN, Alanine, Animaux, Antiviraux, Cellules Sf9, Expression des gènes, Inhibiteurs de la synthèse d'acide nucléique, Réplication virale.
English descriptors
- KwdEn :
- Adenosine Monophosphate (analogs & derivatives), Adenosine Monophosphate (chemistry), Adenosine Monophosphate (pharmacology), Alanine (analogs & derivatives), Alanine (chemistry), Alanine (pharmacology), Animals, Antiviral Agents (chemistry), Antiviral Agents (pharmacology), Coronavirus (enzymology), Ebolavirus (enzymology), Gene Expression, Middle East Respiratory Syndrome Coronavirus (enzymology), Nucleic Acid Synthesis Inhibitors (chemistry), Nucleic Acid Synthesis Inhibitors (pharmacology), RNA, RNA Replicase (antagonists & inhibitors), RNA Replicase (genetics), Sf9 Cells, Viral Nonstructural Proteins (genetics), Virus Replication (drug effects).
- MESH :
- chemical , analogs & derivatives : Adenosine Monophosphate, Alanine.
- chemical , antagonists & inhibitors : RNA Replicase.
- chemical , chemistry : Adenosine Monophosphate, Alanine, Antiviral Agents, Nucleic Acid Synthesis Inhibitors.
- chemical , genetics : RNA Replicase, Viral Nonstructural Proteins.
- chemical , pharmacology : Adenosine Monophosphate, Alanine, Antiviral Agents, Nucleic Acid Synthesis Inhibitors.
- drug effects : Virus Replication.
- enzymology : Coronavirus, Ebolavirus, Middle East Respiratory Syndrome Coronavirus.
- Animals, Gene Expression, RNA, Sf9 Cells.
Abstract
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
Url:
DOI: 10.1074/jbc.AC120.013056
PubMed: 32094225
PubMed Central: 7152756
Affiliations:
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Le document en format XML
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Gordon</name><affiliation wicri:level="1"><nlm:aff id="aff1">Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta T6G 2E1, Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta T6G 2E1</wicri:regionArea><wicri:noRegion>Alberta T6G 2E1</wicri:noRegion></affiliation></author><author><name sortKey="Tchesnokov, Egor P" sort="Tchesnokov, Egor P" uniqKey="Tchesnokov E" first="Egor P." last="Tchesnokov">Egor P. Tchesnokov</name><affiliation wicri:level="1"><nlm:aff id="aff1">Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta T6G 2E1, Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta T6G 2E1</wicri:regionArea><wicri:noRegion>Alberta T6G 2E1</wicri:noRegion></affiliation></author><author><name sortKey="Feng, Joy Y" sort="Feng, Joy Y" uniqKey="Feng J" first="Joy Y." last="Feng">Joy Y. Feng</name><affiliation wicri:level="2"><nlm:aff id="aff2">Gilead Sciences, Inc., Foster City, California 94404</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Californie</region></placeName><wicri:cityArea>Gilead Sciences, Inc., Foster City</wicri:cityArea></affiliation></author><author><name sortKey="Porter, Danielle P" sort="Porter, Danielle P" uniqKey="Porter D" first="Danielle P." last="Porter">Danielle P. Porter</name><affiliation wicri:level="2"><nlm:aff id="aff2">Gilead Sciences, Inc., Foster City, California 94404</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Californie</region></placeName><wicri:cityArea>Gilead Sciences, Inc., Foster City</wicri:cityArea></affiliation></author><author><name sortKey="Gotte, Matthias" sort="Gotte, Matthias" uniqKey="Gotte M" first="Matthias" last="Götte">Matthias Götte</name><affiliation wicri:level="1"><nlm:aff id="aff1">Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta T6G 2E1, Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta T6G 2E1</wicri:regionArea><wicri:noRegion>Alberta T6G 2E1</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff3">Li Ka Shing Institute of Virology at University of Alberta, Edmonton, Alberta T6G 2E1, Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>Li Ka Shing Institute of Virology at University of Alberta, Edmonton, Alberta T6G 2E1</wicri:regionArea><wicri:noRegion>Alberta T6G 2E1</wicri:noRegion></affiliation></author></analytic><series><title level="j">The Journal of Biological Chemistry</title><idno type="ISSN">0021-9258</idno><idno type="eISSN">1083-351X</idno><imprint><date when="2020">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Adenosine Monophosphate (analogs & derivatives)</term><term>Adenosine Monophosphate (chemistry)</term><term>Adenosine Monophosphate (pharmacology)</term><term>Alanine (analogs & derivatives)</term><term>Alanine (chemistry)</term><term>Alanine (pharmacology)</term><term>Animals</term><term>Antiviral Agents (chemistry)</term><term>Antiviral Agents (pharmacology)</term><term>Coronavirus (enzymology)</term><term>Ebolavirus (enzymology)</term><term>Gene Expression</term><term>Middle East Respiratory Syndrome Coronavirus (enzymology)</term><term>Nucleic Acid Synthesis Inhibitors (chemistry)</term><term>Nucleic Acid Synthesis Inhibitors (pharmacology)</term><term>RNA</term><term>RNA Replicase (antagonists & inhibitors)</term><term>RNA Replicase (genetics)</term><term>Sf9 Cells</term><term>Viral Nonstructural Proteins (genetics)</term><term>Virus Replication (drug effects)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>AMP ()</term><term>AMP (analogues et dérivés)</term><term>AMP (pharmacologie)</term><term>ARN</term><term>Alanine ()</term><term>Alanine (analogues et dérivés)</term><term>Alanine (pharmacologie)</term><term>Animaux</term><term>Antiviraux ()</term><term>Antiviraux (pharmacologie)</term><term>Cellules Sf9</term><term>Coronavirus (enzymologie)</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient (enzymologie)</term><term>Ebolavirus (enzymologie)</term><term>Expression des gènes</term><term>Inhibiteurs de la synthèse d'acide nucléique ()</term><term>Inhibiteurs de la synthèse d'acide nucléique (pharmacologie)</term><term>Protéines virales non structurales (génétique)</term><term>RNA replicase (antagonistes et inhibiteurs)</term><term>RNA replicase (génétique)</term><term>Réplication virale ()</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analogs & derivatives" xml:lang="en"><term>Adenosine Monophosphate</term><term>Alanine</term></keywords><keywords scheme="MESH" type="chemical" qualifier="antagonists & inhibitors" xml:lang="en"><term>RNA Replicase</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Adenosine Monophosphate</term><term>Alanine</term><term>Antiviral Agents</term><term>Nucleic Acid Synthesis Inhibitors</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>RNA Replicase</term><term>Viral Nonstructural Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Adenosine Monophosphate</term><term>Alanine</term><term>Antiviral Agents</term><term>Nucleic Acid Synthesis Inhibitors</term></keywords><keywords scheme="MESH" qualifier="analogues et dérivés" xml:lang="fr"><term>AMP</term><term>Alanine</term></keywords><keywords scheme="MESH" qualifier="antagonistes et inhibiteurs" xml:lang="fr"><term>RNA replicase</term></keywords><keywords scheme="MESH" qualifier="drug effects" xml:lang="en"><term>Virus Replication</term></keywords><keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Coronavirus</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient</term><term>Ebolavirus</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Coronavirus</term><term>Ebolavirus</term><term>Middle East Respiratory Syndrome Coronavirus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Protéines virales non structurales</term><term>RNA replicase</term></keywords><keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>AMP</term><term>Alanine</term><term>Antiviraux</term><term>Inhibiteurs de la synthèse d'acide nucléique</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Gene Expression</term><term>RNA</term><term>Sf9 Cells</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>AMP</term><term>ARN</term><term>Alanine</term><term>Animaux</term><term>Antiviraux</term><term>Cellules Sf9</term><term>Expression des gènes</term><term>Inhibiteurs de la synthèse d'acide nucléique</term><term>Réplication virale</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>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 <italic>i</italic>, the inhibitor caused RNA synthesis arrest at position <italic>i</italic> + 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.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Sheahan, T P" uniqKey="Sheahan T">T. P. Sheahan</name></author><author><name sortKey="Sims, A C" uniqKey="Sims A">A. C. Sims</name></author><author><name sortKey="Leist, S R" uniqKey="Leist S">S. R. Leist</name></author><author><name sortKey="Sch Fer, A" uniqKey="Sch Fer A">A. Schäfer</name></author><author><name sortKey="Won, J" uniqKey="Won J">J. Won</name></author><author><name sortKey="Brown, A J" uniqKey="Brown A">A. J. Brown</name></author><author><name sortKey="Montgomery, S A" uniqKey="Montgomery S">S. A. Montgomery</name></author><author><name sortKey="Hogg, A" uniqKey="Hogg A">A. 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Gordon</name></noRegion><name sortKey="Gotte, Matthias" sort="Gotte, Matthias" uniqKey="Gotte M" first="Matthias" last="Götte">Matthias Götte</name><name sortKey="Gotte, Matthias" sort="Gotte, Matthias" uniqKey="Gotte M" first="Matthias" last="Götte">Matthias Götte</name><name sortKey="Tchesnokov, Egor P" sort="Tchesnokov, Egor P" uniqKey="Tchesnokov E" first="Egor P." last="Tchesnokov">Egor P. Tchesnokov</name></country><country name="États-Unis"><region name="Californie"><name sortKey="Feng, Joy Y" sort="Feng, Joy Y" uniqKey="Feng J" first="Joy Y." last="Feng">Joy Y. Feng</name></region><name sortKey="Porter, Danielle P" sort="Porter, Danielle P" uniqKey="Porter D" first="Danielle P." last="Porter">Danielle P. Porter</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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