Inhibitors of foot and mouth disease virus targeting a novel pocket of the RNA-dependent RNA polymerase.
Foot-and-Mouth Disease Virus (FMDV) is a picornavirus that infects cloven-hoofed animals and leads to severe losses in livestock production. In the case of an FMD outbreak, emergency vaccination requires at least 7 days to trigger an effective immune response. There are currently no approved inhibit...
| Main Authors: | , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2010-12-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3006429?pdf=render |
| _version_ | 1818997066526883840 |
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| author | Ryan C Durk Kamalendra Singh Ceili A Cornelison Devendra K Rai Kayla B Matzek Maxwell D Leslie Elizabeth Schafer Bruno Marchand Adeyemi Adedeji Eleftherios Michailidis Christopher A Dorst Jennifer Moran Christie Pautler Luis L Rodriguez Mark A McIntosh Elizabeth Rieder Stefan G Sarafianos |
| author_facet | Ryan C Durk Kamalendra Singh Ceili A Cornelison Devendra K Rai Kayla B Matzek Maxwell D Leslie Elizabeth Schafer Bruno Marchand Adeyemi Adedeji Eleftherios Michailidis Christopher A Dorst Jennifer Moran Christie Pautler Luis L Rodriguez Mark A McIntosh Elizabeth Rieder Stefan G Sarafianos |
| author_sort | Ryan C Durk |
| collection | DOAJ |
| description | Foot-and-Mouth Disease Virus (FMDV) is a picornavirus that infects cloven-hoofed animals and leads to severe losses in livestock production. In the case of an FMD outbreak, emergency vaccination requires at least 7 days to trigger an effective immune response. There are currently no approved inhibitors for the treatment or prevention of FMDV infections.Using a luciferase-based assay we screened a library of compounds and identified seven novel inhibitors of 3Dpol, the RNA-dependent RNA polymerase of FMDV. The compounds inhibited specifically 3Dpol (IC(50)s from 2-17 µM) and not other viral or bacterial polymerases. Enzyme kinetic studies on the inhibition mechanism by compounds 5D9 and 7F8 showed that they are non-competitive inhibitors with respect to NTP and nucleic acid substrates. Molecular modeling and docking studies into the 3Dpol structure revealed an inhibitor binding pocket proximal to, but distinct from the 3Dpol catalytic site. Residues surrounding this pocket are conserved among all 60 FMDV subtypes. Site directed mutagenesis of two residues located at either side of the pocket caused distinct resistance to the compounds, demonstrating that they indeed bind at this site. Several compounds inhibited viral replication with 5D9 suppressing virus production in FMDV-infected cells with EC(50) = 12 µM and EC(90) = 20 µM).We identified several non-competitive inhibitors of FMDV 3Dpol that target a novel binding pocket, which can be used for future structure-based drug design studies. Such studies can lead to the discovery of even more potent antivirals that could provide alternative or supplementary options to contain future outbreaks of FMD. |
| first_indexed | 2024-12-20T21:39:43Z |
| format | Article |
| id | doaj.art-f0122b2af81f4aea9a4ce19a093de893 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-20T21:39:43Z |
| publishDate | 2010-12-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-f0122b2af81f4aea9a4ce19a093de8932022-12-21T19:25:51ZengPublic Library of Science (PLoS)PLoS ONE1932-62032010-12-01512e1504910.1371/journal.pone.0015049Inhibitors of foot and mouth disease virus targeting a novel pocket of the RNA-dependent RNA polymerase.Ryan C DurkKamalendra SinghCeili A CornelisonDevendra K RaiKayla B MatzekMaxwell D LeslieElizabeth SchaferBruno MarchandAdeyemi AdedejiEleftherios MichailidisChristopher A DorstJennifer MoranChristie PautlerLuis L RodriguezMark A McIntoshElizabeth RiederStefan G SarafianosFoot-and-Mouth Disease Virus (FMDV) is a picornavirus that infects cloven-hoofed animals and leads to severe losses in livestock production. In the case of an FMD outbreak, emergency vaccination requires at least 7 days to trigger an effective immune response. There are currently no approved inhibitors for the treatment or prevention of FMDV infections.Using a luciferase-based assay we screened a library of compounds and identified seven novel inhibitors of 3Dpol, the RNA-dependent RNA polymerase of FMDV. The compounds inhibited specifically 3Dpol (IC(50)s from 2-17 µM) and not other viral or bacterial polymerases. Enzyme kinetic studies on the inhibition mechanism by compounds 5D9 and 7F8 showed that they are non-competitive inhibitors with respect to NTP and nucleic acid substrates. Molecular modeling and docking studies into the 3Dpol structure revealed an inhibitor binding pocket proximal to, but distinct from the 3Dpol catalytic site. Residues surrounding this pocket are conserved among all 60 FMDV subtypes. Site directed mutagenesis of two residues located at either side of the pocket caused distinct resistance to the compounds, demonstrating that they indeed bind at this site. Several compounds inhibited viral replication with 5D9 suppressing virus production in FMDV-infected cells with EC(50) = 12 µM and EC(90) = 20 µM).We identified several non-competitive inhibitors of FMDV 3Dpol that target a novel binding pocket, which can be used for future structure-based drug design studies. Such studies can lead to the discovery of even more potent antivirals that could provide alternative or supplementary options to contain future outbreaks of FMD.http://europepmc.org/articles/PMC3006429?pdf=render |
| spellingShingle | Ryan C Durk Kamalendra Singh Ceili A Cornelison Devendra K Rai Kayla B Matzek Maxwell D Leslie Elizabeth Schafer Bruno Marchand Adeyemi Adedeji Eleftherios Michailidis Christopher A Dorst Jennifer Moran Christie Pautler Luis L Rodriguez Mark A McIntosh Elizabeth Rieder Stefan G Sarafianos Inhibitors of foot and mouth disease virus targeting a novel pocket of the RNA-dependent RNA polymerase. PLoS ONE |
| title | Inhibitors of foot and mouth disease virus targeting a novel pocket of the RNA-dependent RNA polymerase. |
| title_full | Inhibitors of foot and mouth disease virus targeting a novel pocket of the RNA-dependent RNA polymerase. |
| title_fullStr | Inhibitors of foot and mouth disease virus targeting a novel pocket of the RNA-dependent RNA polymerase. |
| title_full_unstemmed | Inhibitors of foot and mouth disease virus targeting a novel pocket of the RNA-dependent RNA polymerase. |
| title_short | Inhibitors of foot and mouth disease virus targeting a novel pocket of the RNA-dependent RNA polymerase. |
| title_sort | inhibitors of foot and mouth disease virus targeting a novel pocket of the rna dependent rna polymerase |
| url | http://europepmc.org/articles/PMC3006429?pdf=render |
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