SrasV1, PubMed, Checkpoint, bibRecord, 001555

Zn(2+) inhibits coronavirus and arterivirus RNA polymerase activity in vitro and zinc ionophores block the replication of these viruses in cell culture.

Identifieur interne : 001555 ( PubMed/Checkpoint ); précédent : 001554; suivant : 001556

Zn(2+) inhibits coronavirus and arterivirus RNA polymerase activity in vitro and zinc ionophores block the replication of these viruses in cell culture.

Auteurs : Aartjan J W. Te Velthuis [Pays-Bas] ; Sjoerd H E. Van Den Worm ; Amy C. Sims ; Ralph S. Baric ; Eric J. Snijder ; Martijn J. Van Hemert

Source :

PLoS pathogens [ 1553-7374 ] ; 2010.

RBID : pubmed:21079686

Descripteurs français

KwdFr :
- ARN messager (génétique), ARN viral (génétique), ARN viral (métabolisme), Animaux, Arterivirus (), Arterivirus (enzymologie), Cellules Vero, Composés du zinc (pharmacologie), Escherichia coli (enzymologie), Escherichia coli (génétique), Infections à artérivirus (anatomopathologie), Infections à artérivirus (traitement médicamenteux), Infections à artérivirus (virologie), Ionophores (pharmacologie), RNA replicase (antagonistes et inhibiteurs), RNA replicase (métabolisme), RT-PCR, Réplication virale (), Syndrome respiratoire aigu sévère (anatomopathologie), Syndrome respiratoire aigu sévère (traitement médicamenteux), Syndrome respiratoire aigu sévère (virologie), Technique de Western, Techniques in vitro, Test de retard de migration électrophorétique, Virus du SRAS (), Virus du SRAS (enzymologie).
MESH :
- anatomopathologie : Infections à artérivirus, Syndrome respiratoire aigu sévère.
- antagonistes et inhibiteurs : RNA replicase.
- enzymologie : Arterivirus, Escherichia coli, Virus du SRAS.
- génétique : ARN messager, ARN viral, Escherichia coli.
- métabolisme : ARN viral, RNA replicase.
- pharmacologie : Composés du zinc, Ionophores.
- traitement médicamenteux : Infections à artérivirus, Syndrome respiratoire aigu sévère.
- virologie : Infections à artérivirus, Syndrome respiratoire aigu sévère.
- Animaux, Arterivirus, Cellules Vero, RT-PCR, Réplication virale, Technique de Western, Techniques in vitro, Test de retard de migration électrophorétique, Virus du SRAS.

English descriptors

KwdEn :
- Animals, Arterivirus (drug effects), Arterivirus (enzymology), Arterivirus Infections (drug therapy), Arterivirus Infections (pathology), Arterivirus Infections (virology), Blotting, Western, Chlorocebus aethiops, Electrophoretic Mobility Shift Assay, Escherichia coli (enzymology), Escherichia coli (genetics), In Vitro Techniques, Ionophores (pharmacology), RNA Replicase (antagonists & inhibitors), RNA Replicase (metabolism), RNA, Messenger (genetics), RNA, Viral (genetics), RNA, Viral (metabolism), Reverse Transcriptase Polymerase Chain Reaction, SARS Virus (drug effects), SARS Virus (enzymology), Severe Acute Respiratory Syndrome (drug therapy), Severe Acute Respiratory Syndrome (pathology), Severe Acute Respiratory Syndrome (virology), Vero Cells, Virus Replication (drug effects), Zinc Compounds (pharmacology).
MESH :
- chemical , antagonists & inhibitors : RNA Replicase.
- chemical , genetics : RNA, Messenger, RNA, Viral.
- chemical , metabolism : RNA Replicase, RNA, Viral.
- chemical , pharmacology : Ionophores, Zinc Compounds.
- drug effects : Arterivirus, SARS Virus, Virus Replication.
- drug therapy : Arterivirus Infections, Severe Acute Respiratory Syndrome.
- enzymology : Arterivirus, Escherichia coli, SARS Virus.
- genetics : Escherichia coli.
- pathology : Arterivirus Infections, Severe Acute Respiratory Syndrome.
- virology : Arterivirus Infections, Severe Acute Respiratory Syndrome.
- Animals, Blotting, Western, Chlorocebus aethiops, Electrophoretic Mobility Shift Assay, In Vitro Techniques, Reverse Transcriptase Polymerase Chain Reaction, Vero Cells.

Abstract

Increasing the intracellular Zn(2+) concentration with zinc-ionophores like pyrithione (PT) can efficiently impair the replication of a variety of RNA viruses, including poliovirus and influenza virus. For some viruses this effect has been attributed to interference with viral polyprotein processing. In this study we demonstrate that the combination of Zn(2+) and PT at low concentrations (2 µM Zn(2+) and 2 µM PT) inhibits the replication of SARS-coronavirus (SARS-CoV) and equine arteritis virus (EAV) in cell culture. The RNA synthesis of these two distantly related nidoviruses is catalyzed by an RNA-dependent RNA polymerase (RdRp), which is the core enzyme of their multiprotein replication and transcription complex (RTC). Using an activity assay for RTCs isolated from cells infected with SARS-CoV or EAV--thus eliminating the need for PT to transport Zn(2+) across the plasma membrane--we show that Zn(2+) efficiently inhibits the RNA-synthesizing activity of the RTCs of both viruses. Enzymatic studies using recombinant RdRps (SARS-CoV nsp12 and EAV nsp9) purified from E. coli subsequently revealed that Zn(2+) directly inhibited the in vitro activity of both nidovirus polymerases. More specifically, Zn(2+) was found to block the initiation step of EAV RNA synthesis, whereas in the case of the SARS-CoV RdRp elongation was inhibited and template binding reduced. By chelating Zn(2+) with MgEDTA, the inhibitory effect of the divalent cation could be reversed, which provides a novel experimental tool for in vitro studies of the molecular details of nidovirus replication and transcription.

DOI: 10.1371/journal.ppat.1001176
PubMed: 21079686

Affiliations:

Links toward previous steps (curation, corpus...)

to stream PubMed, to step Corpus: 001587
to stream PubMed, to step Curation: 001587

Links to Exploration step

pubmed:21079686

Le document en format XML

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<front><div type="abstract" xml:lang="en">Increasing the intracellular Zn(2+) concentration with zinc-ionophores like pyrithione (PT) can efficiently impair the replication of a variety of RNA viruses, including poliovirus and influenza virus. For some viruses this effect has been attributed to interference with viral polyprotein processing. In this study we demonstrate that the combination of Zn(2+) and PT at low concentrations (2 µM Zn(2+) and 2 µM PT) inhibits the replication of SARS-coronavirus (SARS-CoV) and equine arteritis virus (EAV) in cell culture. The RNA synthesis of these two distantly related nidoviruses is catalyzed by an RNA-dependent RNA polymerase (RdRp), which is the core enzyme of their multiprotein replication and transcription complex (RTC). Using an activity assay for RTCs isolated from cells infected with SARS-CoV or EAV--thus eliminating the need for PT to transport Zn(2+) across the plasma membrane--we show that Zn(2+) efficiently inhibits the RNA-synthesizing activity of the RTCs of both viruses. Enzymatic studies using recombinant RdRps (SARS-CoV nsp12 and EAV nsp9) purified from E. coli subsequently revealed that Zn(2+) directly inhibited the in vitro activity of both nidovirus polymerases. More specifically, Zn(2+) was found to block the initiation step of EAV RNA synthesis, whereas in the case of the SARS-CoV RdRp elongation was inhibited and template binding reduced. By chelating Zn(2+) with MgEDTA, the inhibitory effect of the divalent cation could be reversed, which provides a novel experimental tool for in vitro studies of the molecular details of nidovirus replication and transcription.</div>
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<Abstract><AbstractText>Increasing the intracellular Zn(2+) concentration with zinc-ionophores like pyrithione (PT) can efficiently impair the replication of a variety of RNA viruses, including poliovirus and influenza virus. For some viruses this effect has been attributed to interference with viral polyprotein processing. In this study we demonstrate that the combination of Zn(2+) and PT at low concentrations (2 µM Zn(2+) and 2 µM PT) inhibits the replication of SARS-coronavirus (SARS-CoV) and equine arteritis virus (EAV) in cell culture. The RNA synthesis of these two distantly related nidoviruses is catalyzed by an RNA-dependent RNA polymerase (RdRp), which is the core enzyme of their multiprotein replication and transcription complex (RTC). Using an activity assay for RTCs isolated from cells infected with SARS-CoV or EAV--thus eliminating the need for PT to transport Zn(2+) across the plasma membrane--we show that Zn(2+) efficiently inhibits the RNA-synthesizing activity of the RTCs of both viruses. Enzymatic studies using recombinant RdRps (SARS-CoV nsp12 and EAV nsp9) purified from E. coli subsequently revealed that Zn(2+) directly inhibited the in vitro activity of both nidovirus polymerases. More specifically, Zn(2+) was found to block the initiation step of EAV RNA synthesis, whereas in the case of the SARS-CoV RdRp elongation was inhibited and template binding reduced. By chelating Zn(2+) with MgEDTA, the inhibitory effect of the divalent cation could be reversed, which provides a novel experimental tool for in vitro studies of the molecular details of nidovirus replication and transcription.</AbstractText>
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Zn(2+) inhibits coronavirus and arterivirus RNA polymerase activity in vitro and zinc ionophores block the replication of these viruses in cell culture.

Zn(2+) inhibits coronavirus and arterivirus RNA polymerase activity in vitro and zinc ionophores block the replication of these viruses in cell culture.

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