Zn(2+) inhibits coronavirus and arterivirus RNA polymerase activity in vitro and zinc ionophores block the replication of these viruses in cell culture
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...
| Main Authors: | , , , , , |
|---|---|
| Other Authors: | |
| Format: | Journal article |
| Language: | English |
| Published: |
2010
|
| _version_ | 1797086743370924032 |
|---|---|
| author | Te Velthuis, A van den Worm, S Sims, A Baric, R Snijder, E van Hemert, M |
| author2 | Andino, R |
| author_facet | Andino, R Te Velthuis, A van den Worm, S Sims, A Baric, R Snijder, E van Hemert, M |
| author_sort | Te Velthuis, A |
| collection | OXFORD |
| description | 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 |
| first_indexed | 2024-03-07T02:26:19Z |
| format | Journal article |
| id | oxford-uuid:a5bc896b-01d1-45aa-850e-72089b7c7fee |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T02:26:19Z |
| publishDate | 2010 |
| record_format | dspace |
| spelling | oxford-uuid:a5bc896b-01d1-45aa-850e-72089b7c7fee2022-03-27T02:42:30ZZn(2+) inhibits coronavirus and arterivirus RNA polymerase activity in vitro and zinc ionophores block the replication of these viruses in cell cultureJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:a5bc896b-01d1-45aa-850e-72089b7c7feeEnglishSymplectic Elements at Oxford2010Te Velthuis, Avan den Worm, SSims, ABaric, RSnijder, Evan Hemert, MAndino, RIncreasing 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 |
| spellingShingle | Te Velthuis, A van den Worm, S Sims, A Baric, R Snijder, E van Hemert, M Zn(2+) inhibits coronavirus and arterivirus RNA polymerase activity in vitro and zinc ionophores block the replication of these viruses in cell culture |
| title | Zn(2+) inhibits coronavirus and arterivirus RNA polymerase activity in vitro and zinc ionophores block the replication of these viruses in cell culture |
| title_full | Zn(2+) inhibits coronavirus and arterivirus RNA polymerase activity in vitro and zinc ionophores block the replication of these viruses in cell culture |
| title_fullStr | Zn(2+) inhibits coronavirus and arterivirus RNA polymerase activity in vitro and zinc ionophores block the replication of these viruses in cell culture |
| title_full_unstemmed | Zn(2+) inhibits coronavirus and arterivirus RNA polymerase activity in vitro and zinc ionophores block the replication of these viruses in cell culture |
| title_short | Zn(2+) inhibits coronavirus and arterivirus RNA polymerase activity in vitro and zinc ionophores block the replication of these viruses in cell culture |
| title_sort | zn 2 inhibits coronavirus and arterivirus rna polymerase activity in vitro and zinc ionophores block the replication of these viruses in cell culture |
| work_keys_str_mv | AT tevelthuisa zn2inhibitscoronavirusandarterivirusrnapolymeraseactivityinvitroandzincionophoresblockthereplicationofthesevirusesincellculture AT vandenworms zn2inhibitscoronavirusandarterivirusrnapolymeraseactivityinvitroandzincionophoresblockthereplicationofthesevirusesincellculture AT simsa zn2inhibitscoronavirusandarterivirusrnapolymeraseactivityinvitroandzincionophoresblockthereplicationofthesevirusesincellculture AT baricr zn2inhibitscoronavirusandarterivirusrnapolymeraseactivityinvitroandzincionophoresblockthereplicationofthesevirusesincellculture AT snijdere zn2inhibitscoronavirusandarterivirusrnapolymeraseactivityinvitroandzincionophoresblockthereplicationofthesevirusesincellculture AT vanhemertm zn2inhibitscoronavirusandarterivirusrnapolymeraseactivityinvitroandzincionophoresblockthereplicationofthesevirusesincellculture |