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Targeting intramolecular proteinase NS2B/3 cleavages for trans-dominant inhibition of dengue virus

Many positive-strand RNA viruses translate their genomes as single polyproteins that are processed by host and viral proteinases to generate all viral protein products. Among these is dengue virus, which encodes the serine proteinase NS2B/3 responsible for seven different cleavages in the polyprotei...

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Autores principales: Constant, David A., Mateo, Roberto, Nagamine, Claude M., Kirkegaard, Karla
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6176606/
https://www.ncbi.nlm.nih.gov/pubmed/30228122
http://dx.doi.org/10.1073/pnas.1805195115
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author Constant, David A.
Mateo, Roberto
Nagamine, Claude M.
Kirkegaard, Karla
author_facet Constant, David A.
Mateo, Roberto
Nagamine, Claude M.
Kirkegaard, Karla
author_sort Constant, David A.
collection PubMed
description Many positive-strand RNA viruses translate their genomes as single polyproteins that are processed by host and viral proteinases to generate all viral protein products. Among these is dengue virus, which encodes the serine proteinase NS2B/3 responsible for seven different cleavages in the polyprotein. NS2B/3 has been the subject of many directed screens to find chemical inhibitors, of which the compound ARDP0006 is among the most effective at inhibiting viral growth. We show that at least three cleavages in the dengue polyprotein are exclusively intramolecular. By definition, such a cis-acting defect cannot be rescued in trans. This creates the possibility that a drug-susceptible or inhibited proteinase can be genetically dominant, inhibiting the outgrowth of drug-resistant virus via precursor accumulation. Indeed, an NS3-G459L variant that is incapable of cleavage at the internal NS3 junction dominantly inhibited negative-strand RNA synthesis of wild-type virus present in the same cell. This internal NS3 cleavage site is the junction most inhibited by ARDP0006, making it likely that the accumulation of toxic precursors, not inhibition of proteolytic activity per se, explains the antiviral efficacy of this compound in restraining viral growth. We argue that intramolecularly cleaving proteinases are promising drug targets for viruses that encode polyproteins. The most effective inhibitors will specifically target cleavage sites required for processing precursors that exert trans-dominant inhibition.
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spelling pubmed-61766062018-10-11 Targeting intramolecular proteinase NS2B/3 cleavages for trans-dominant inhibition of dengue virus Constant, David A. Mateo, Roberto Nagamine, Claude M. Kirkegaard, Karla Proc Natl Acad Sci U S A Biological Sciences Many positive-strand RNA viruses translate their genomes as single polyproteins that are processed by host and viral proteinases to generate all viral protein products. Among these is dengue virus, which encodes the serine proteinase NS2B/3 responsible for seven different cleavages in the polyprotein. NS2B/3 has been the subject of many directed screens to find chemical inhibitors, of which the compound ARDP0006 is among the most effective at inhibiting viral growth. We show that at least three cleavages in the dengue polyprotein are exclusively intramolecular. By definition, such a cis-acting defect cannot be rescued in trans. This creates the possibility that a drug-susceptible or inhibited proteinase can be genetically dominant, inhibiting the outgrowth of drug-resistant virus via precursor accumulation. Indeed, an NS3-G459L variant that is incapable of cleavage at the internal NS3 junction dominantly inhibited negative-strand RNA synthesis of wild-type virus present in the same cell. This internal NS3 cleavage site is the junction most inhibited by ARDP0006, making it likely that the accumulation of toxic precursors, not inhibition of proteolytic activity per se, explains the antiviral efficacy of this compound in restraining viral growth. We argue that intramolecularly cleaving proteinases are promising drug targets for viruses that encode polyproteins. The most effective inhibitors will specifically target cleavage sites required for processing precursors that exert trans-dominant inhibition. National Academy of Sciences 2018-10-02 2018-09-18 /pmc/articles/PMC6176606/ /pubmed/30228122 http://dx.doi.org/10.1073/pnas.1805195115 Text en Copyright © 2018 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/ This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) .
spellingShingle Biological Sciences
Constant, David A.
Mateo, Roberto
Nagamine, Claude M.
Kirkegaard, Karla
Targeting intramolecular proteinase NS2B/3 cleavages for trans-dominant inhibition of dengue virus
title Targeting intramolecular proteinase NS2B/3 cleavages for trans-dominant inhibition of dengue virus
title_full Targeting intramolecular proteinase NS2B/3 cleavages for trans-dominant inhibition of dengue virus
title_fullStr Targeting intramolecular proteinase NS2B/3 cleavages for trans-dominant inhibition of dengue virus
title_full_unstemmed Targeting intramolecular proteinase NS2B/3 cleavages for trans-dominant inhibition of dengue virus
title_short Targeting intramolecular proteinase NS2B/3 cleavages for trans-dominant inhibition of dengue virus
title_sort targeting intramolecular proteinase ns2b/3 cleavages for trans-dominant inhibition of dengue virus
topic Biological Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6176606/
https://www.ncbi.nlm.nih.gov/pubmed/30228122
http://dx.doi.org/10.1073/pnas.1805195115
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