Show simple item record

dc.contributor.authorDultz, Georg
dc.contributor.authorSrikakulam, Sanjay K
dc.contributor.authorKonetschnik, Michael
dc.contributor.authorShimakami, Tetsuro
dc.contributor.authorDoncheva, Nadezhda T
dc.contributor.authorDietz, Julia
dc.contributor.authorSarrazin, Christoph
dc.contributor.authorBiondi, Ricardo M
dc.contributor.authorZeuzem, Stefan
dc.contributor.authorTampé, Robert
dc.contributor.authorKalinina, Olga V
dc.contributor.authorWelsch, Christoph
dc.date.accessioned2021-10-29T14:31:07Z
dc.date.available2021-10-29T14:31:07Z
dc.date.issued2021-07-31
dc.identifier.citationBiol Chem. 2021 Sep;297(3):101031. doi: 10.1016/j.jbc.2021.101031. Epub 2021 Jul 31.en_US
dc.identifier.pmid34339738
dc.identifier.doi10.1016/j.jbc.2021.101031
dc.identifier.urihttp://hdl.handle.net/10033/623087
dc.description.abstractThe Q80K polymorphism in the NS3-4A protease of the hepatitis C virus is associated with treatment failure of direct-acting antiviral agents. This polymorphism is highly prevalent in genotype 1a infections and stably transmitted between hosts. Here, we investigated the underlying molecular mechanisms of evolutionarily conserved coevolving amino acids in NS3-Q80K and revealed potential implications of epistatic interactions in immune escape and variants persistence. Using purified protein, we characterized the impact of epistatic amino acid substitutions on the physicochemical properties and peptide cleavage kinetics of the NS3-Q80K protease. We found that Q80K destabilized the protease protein fold (p < 0.0001). Although NS3-Q80K showed reduced peptide substrate turnover (p < 0.0002), replicative fitness in an H77S.3 cell culture model of infection was not significantly inferior to the WT virus. Epistatic substitutions at residues 91 and 174 in NS3-Q80K stabilized the protein fold (p < 0.0001) and leveraged the WT protease stability. However, changes in protease stability inversely correlated with enzymatic activity. In infectious cell culture, these secondary substitutions were not associated with a gain of replicative fitness in NS3-Q80K variants. Using molecular dynamics, we observed that the total number of residue contacts in NS3-Q80K mutants correlated with protein folding stability. Changes in the number of contacts reflected the compensatory effect on protein folding instability by epistatic substitutions. In summary, epistatic substitutions in NS3-Q80K contribute to viral fitness by mechanisms not directly related to RNA replication. By compensating for protein-folding instability, epistatic interactions likely protect NS3-Q80K variants from immune cell recognition.en_US
dc.language.isoenen_US
dc.publisherElsevieren_US
dc.rightsAttribution 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/*
dc.subjecthepatitis C virusen_US
dc.subjectimmune escapeen_US
dc.subjectprotein evolutionen_US
dc.subjectprotein foldingen_US
dc.subjectresistance mutationen_US
dc.subjectserine protease (NS3-4A)en_US
dc.subjectviral fitnessen_US
dc.titleEpistatic interactions promote persistence of NS3-Q80K in HCV infection by compensating for protein folding instability.en_US
dc.typeArticleen_US
dc.identifier.eissn1083-351X
dc.contributor.departmentHIPS, Helmholtz-Institut für Pharmazeutische Forschung Saarland, Universitätscampus E8.1 66123 Saarbrücken, Germany.en_US
dc.identifier.journalThe Journal of biological chemistryen_US
dc.source.volume297
dc.source.issue3
dc.source.beginpage101031
dc.source.endpage
refterms.dateFOA2021-10-29T14:31:08Z
dc.source.journaltitleThe Journal of biological chemistry
dc.source.countryUnited States


Files in this item

Thumbnail
Name:
Publisher version
Thumbnail
Name:
Dultz et al.pdf
Size:
1.354Mb
Format:
PDF
Description:
Open Access publication

This item appears in the following Collection(s)

Show simple item record

Attribution 4.0 International
Except where otherwise noted, this item's license is described as Attribution 4.0 International