Please use this identifier to cite or link to this item:
https://doi.org/10.1371/journal.ppat.1005737
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dc.title | Potent Allosteric Dengue Virus NS5 Polymerase Inhibitors: Mechanism of Action and Resistance Profiling | |
dc.contributor.author | Lim S.P. | |
dc.contributor.author | Noble C.G. | |
dc.contributor.author | Seh C.C. | |
dc.contributor.author | Soh T.S. | |
dc.contributor.author | El Sahili A. | |
dc.contributor.author | Chan G.K.Y. | |
dc.contributor.author | Lescar J. | |
dc.contributor.author | Arora R. | |
dc.contributor.author | Benson T. | |
dc.contributor.author | Nilar S. | |
dc.contributor.author | Manjunatha U. | |
dc.contributor.author | Wan K.F. | |
dc.contributor.author | Dong H. | |
dc.contributor.author | Xie X. | |
dc.contributor.author | Shi P.-Y. | |
dc.contributor.author | Yokokawa F. | |
dc.date.accessioned | 2019-11-08T06:48:02Z | |
dc.date.available | 2019-11-08T06:48:02Z | |
dc.date.issued | 2016 | |
dc.identifier.citation | Lim S.P., Noble C.G., Seh C.C., Soh T.S., El Sahili A., Chan G.K.Y., Lescar J., Arora R., Benson T., Nilar S., Manjunatha U., Wan K.F., Dong H., Xie X., Shi P.-Y., Yokokawa F. (2016). Potent Allosteric Dengue Virus NS5 Polymerase Inhibitors: Mechanism of Action and Resistance Profiling. PLoS Pathogens 12 (8) : e1005737. ScholarBank@NUS Repository. https://doi.org/10.1371/journal.ppat.1005737 | |
dc.identifier.issn | 15537366 | |
dc.identifier.uri | https://scholarbank.nus.edu.sg/handle/10635/161910 | |
dc.description.abstract | Flaviviruses comprise major emerging pathogens such as dengue virus (DENV) or Zika virus (ZIKV). The flavivirus RNA genome is replicated by the RNA-dependent-RNA polymerase (RdRp) domain of non-structural protein 5 (NS5). This essential enzymatic activity renders the RdRp attractive for antiviral therapy. NS5 synthesizes viral RNA via a ?de novo? initiation mechanism. Crystal structures of the flavivirus RdRp revealed a ?closed? conformation reminiscent of a pre-initiation state, with a well ordered priming loop that extrudes from the thumb subdomain into the dsRNA exit tunnel, close to the ?GDD? active site. To-date, no allosteric pockets have been identified for the RdRp, and compound screening campaigns did not yield suitable drug candidates. Using fragment-based screening via X-ray crystallography, we found a fragment that bound to a pocket of the apo-DENV RdRp close to its active site (termed ?N pocket?). Structure-guided improvements yielded DENV pan-serotype inhibitors of the RdRp de novo initiation activity with nano-molar potency that also impeded elongation activity at micro-molar concentrations. Inhibitors exhibited mixed inhibition kinetics with respect to competition with the RNA or GTP substrate. The best compounds have EC50 values of 1?2 ?M against all four DENV serotypes in cell culture assays. Genome-sequencing of compound-resistant DENV replicons, identified amino acid changes that mapped to the N pocket. Since inhibitors bind at the thumb/palm interface of the RdRp, this class of compounds is proposed to hinder RdRp conformational changes during its transition from initiation to elongation. This is the first report of a class of pan-serotype and cell-active DENV RdRp inhibitors. Given the evolutionary conservation of residues lining the N pocket, these molecules offer insights to treat other serious conditions caused by flaviviruses. ? 2016 Lim et al. | |
dc.rights | Attribution 4.0 International | |
dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | |
dc.source | Unpaywall 20191101 | |
dc.subject | nonstructural protein 5 | |
dc.subject | RNA directed RNA polymerase | |
dc.subject | antivirus agent | |
dc.subject | NS5 protein, dengue virus | |
dc.subject | nucleic acid synthesis inhibitor | |
dc.subject | RNA directed RNA polymerase | |
dc.subject | viral protein | |
dc.subject | allosterism | |
dc.subject | antiviral therapy | |
dc.subject | Article | |
dc.subject | binding affinity | |
dc.subject | biochemical analysis | |
dc.subject | CC50 | |
dc.subject | controlled study | |
dc.subject | Dengue virus | |
dc.subject | EC50 | |
dc.subject | enzyme activity | |
dc.subject | enzyme assay | |
dc.subject | enzyme inhibition assay | |
dc.subject | Flavivirus | |
dc.subject | gene expression | |
dc.subject | IC50 | |
dc.subject | luciferase assay | |
dc.subject | nonhuman | |
dc.subject | protein expression | |
dc.subject | reverse genetics | |
dc.subject | virus morphology | |
dc.subject | virus replication | |
dc.subject | X ray crystallography | |
dc.subject | Zika virus | |
dc.subject | A-549 cell line | |
dc.subject | antagonists and inhibitors | |
dc.subject | chemistry | |
dc.subject | dengue | |
dc.subject | Dengue virus | |
dc.subject | drug design | |
dc.subject | drug effects | |
dc.subject | human | |
dc.subject | protein domain | |
dc.subject | surface plasmon resonance | |
dc.subject | A549 Cells | |
dc.subject | Antiviral Agents | |
dc.subject | Crystallography, X-Ray | |
dc.subject | Dengue | |
dc.subject | Dengue Virus | |
dc.subject | Drug Design | |
dc.subject | Humans | |
dc.subject | Nucleic Acid Synthesis Inhibitors | |
dc.subject | Protein Domains | |
dc.subject | RNA Replicase | |
dc.subject | Surface Plasmon Resonance | |
dc.subject | Viral Nonstructural Proteins | |
dc.type | Article | |
dc.contributor.department | DEAN'S OFFICE (DUKE-NUS MEDICAL SCHOOL) | |
dc.contributor.department | DUKE-NUS MEDICAL SCHOOL | |
dc.contributor.department | BIOLOGICAL SCIENCES | |
dc.contributor.department | MICROBIOLOGY AND IMMUNOLOGY | |
dc.description.doi | 10.1371/journal.ppat.1005737 | |
dc.description.sourcetitle | PLoS Pathogens | |
dc.description.volume | 12 | |
dc.description.issue | 8 | |
dc.description.page | e1005737 | |
Appears in Collections: | Elements Staff Publications |
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