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Poxviruses Bearing DNA Polymerase Mutations Show Complex Patterns of Cross-Resistance

Despite the eradication of smallpox four decades ago, poxviruses continue to be a threat to humans and animals. The arsenal of anti-poxvirus agents is very limited and understanding mechanisms of resistance to agents targeting viral DNA polymerases is fundamental for the development of antiviral the...

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Autores principales: Andrei, Graciela, Fiten, Pierre, Krečmerová, Marcela, Opdenakker, Ghislain, Topalis, Dimitrios, Snoeck, Robert
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8945813/
https://www.ncbi.nlm.nih.gov/pubmed/35327382
http://dx.doi.org/10.3390/biomedicines10030580
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author Andrei, Graciela
Fiten, Pierre
Krečmerová, Marcela
Opdenakker, Ghislain
Topalis, Dimitrios
Snoeck, Robert
author_facet Andrei, Graciela
Fiten, Pierre
Krečmerová, Marcela
Opdenakker, Ghislain
Topalis, Dimitrios
Snoeck, Robert
author_sort Andrei, Graciela
collection PubMed
description Despite the eradication of smallpox four decades ago, poxviruses continue to be a threat to humans and animals. The arsenal of anti-poxvirus agents is very limited and understanding mechanisms of resistance to agents targeting viral DNA polymerases is fundamental for the development of antiviral therapies. We describe here the phenotypic and genotypic characterization of poxvirus DNA polymerase mutants isolated under selective pressure with different acyclic nucleoside phosphonates, including HPMPC (cidofovir), cHPMPC, HPMPA, cHPMPA, HPMPDAP, HPMPO-DAPy, and PMEO-DAPy, and the pyrophosphate analogue phosphonoacetic acid. Vaccinia virus (VACV) and cowpox virus drug-resistant viral clones emerging under drug pressure were characterized phenotypically (drug-susceptibility profile) and genotypically (DNA polymerase sequencing). Different amino acid changes in the polymerase domain and in the 3′-5′ exonuclease domain were linked to drug resistance. Changes in the 3′-5′ domain emerged earlier than in the polymerase domain when viruses acquired a combination of mutations. Our study highlights the importance of poxvirus DNA polymerase residues 314, 613, 684, 688, and 851, previously linked to drug resistance, and identified several novel mutations in the 3′-5′ exonuclease domain (M313I, F354L, D480Y) and in the DNA polymerase domain (A632T, T831I, E856K, L924F) associated with different drug-susceptibility profiles. Furthermore, a combination of mutations resulted in complex patterns of cross-resistance. Modeling of the VACV DNA polymerase bearing the newly described mutations was performed to understand the effects of these mutations on the structure of the viral enzyme. We demonstrated the emergence of drug-resistant DNA polymerase mutations in complex patterns to be considered in case such mutations should eventually arise in the clinic.
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spelling pubmed-89458132022-03-25 Poxviruses Bearing DNA Polymerase Mutations Show Complex Patterns of Cross-Resistance Andrei, Graciela Fiten, Pierre Krečmerová, Marcela Opdenakker, Ghislain Topalis, Dimitrios Snoeck, Robert Biomedicines Article Despite the eradication of smallpox four decades ago, poxviruses continue to be a threat to humans and animals. The arsenal of anti-poxvirus agents is very limited and understanding mechanisms of resistance to agents targeting viral DNA polymerases is fundamental for the development of antiviral therapies. We describe here the phenotypic and genotypic characterization of poxvirus DNA polymerase mutants isolated under selective pressure with different acyclic nucleoside phosphonates, including HPMPC (cidofovir), cHPMPC, HPMPA, cHPMPA, HPMPDAP, HPMPO-DAPy, and PMEO-DAPy, and the pyrophosphate analogue phosphonoacetic acid. Vaccinia virus (VACV) and cowpox virus drug-resistant viral clones emerging under drug pressure were characterized phenotypically (drug-susceptibility profile) and genotypically (DNA polymerase sequencing). Different amino acid changes in the polymerase domain and in the 3′-5′ exonuclease domain were linked to drug resistance. Changes in the 3′-5′ domain emerged earlier than in the polymerase domain when viruses acquired a combination of mutations. Our study highlights the importance of poxvirus DNA polymerase residues 314, 613, 684, 688, and 851, previously linked to drug resistance, and identified several novel mutations in the 3′-5′ exonuclease domain (M313I, F354L, D480Y) and in the DNA polymerase domain (A632T, T831I, E856K, L924F) associated with different drug-susceptibility profiles. Furthermore, a combination of mutations resulted in complex patterns of cross-resistance. Modeling of the VACV DNA polymerase bearing the newly described mutations was performed to understand the effects of these mutations on the structure of the viral enzyme. We demonstrated the emergence of drug-resistant DNA polymerase mutations in complex patterns to be considered in case such mutations should eventually arise in the clinic. MDPI 2022-03-01 /pmc/articles/PMC8945813/ /pubmed/35327382 http://dx.doi.org/10.3390/biomedicines10030580 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Andrei, Graciela
Fiten, Pierre
Krečmerová, Marcela
Opdenakker, Ghislain
Topalis, Dimitrios
Snoeck, Robert
Poxviruses Bearing DNA Polymerase Mutations Show Complex Patterns of Cross-Resistance
title Poxviruses Bearing DNA Polymerase Mutations Show Complex Patterns of Cross-Resistance
title_full Poxviruses Bearing DNA Polymerase Mutations Show Complex Patterns of Cross-Resistance
title_fullStr Poxviruses Bearing DNA Polymerase Mutations Show Complex Patterns of Cross-Resistance
title_full_unstemmed Poxviruses Bearing DNA Polymerase Mutations Show Complex Patterns of Cross-Resistance
title_short Poxviruses Bearing DNA Polymerase Mutations Show Complex Patterns of Cross-Resistance
title_sort poxviruses bearing dna polymerase mutations show complex patterns of cross-resistance
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8945813/
https://www.ncbi.nlm.nih.gov/pubmed/35327382
http://dx.doi.org/10.3390/biomedicines10030580
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