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dc.contributor.authorKitano, Mitsutaka
dc.contributor.authorHosmillo, Myra
dc.contributor.authorEmmott, Edward
dc.contributor.authorLu, Jia
dc.contributor.authorGoodfellow, Ian
dc.date.accessioned2018-05-18T11:11:27Z
dc.date.available2018-05-18T11:11:27Z
dc.date.issued2018-05
dc.identifier.issn0066-4804
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/275924
dc.description.abstractHuman norovirus (HuNoV) is a major cause of nonbacterial gastroenteritis worldwide, yet despite its impact on society, vaccines and antivirals are currently lacking. A HuNoV replicon system has been widely applied to the evaluation of antiviral compounds and has thus accelerated the process of drug discovery against HuNoV infection. Rupintrivir, an irreversible inhibitor of the human rhinovirus 3C protease, has been reported to inhibit the replication of the Norwalk virus replicon via the inhibition of the norovirus protease. Here we report, for the first time, the generation of rupintrivir-resistant human Norwalk virus replicon cells in vitro Sequence analysis revealed that these replicon cells contained amino acid substitutions of alanine 105 to valine (A105V) and isoleucine 109 to valine (I109V) in the viral protease NS6. The application of a cell-based fluorescence resonance energy transfer (FRET) assay for protease activity demonstrated that these substitutions were involved in the enhanced resistance to rupintrivir. Furthermore, we validated the effect of these mutations using reverse genetics in murine norovirus (MNV), demonstrating that a recombinant MNV strain with a single I109V substitution in the protease also showed reduced susceptibility to rupintrivir. In summary, using a combination of different approaches, we have demonstrated that, under the correct conditions, mutations in the norovirus protease that lead to the generation of resistant mutants can rapidly occur.
dc.format.mediumElectronic-Print
dc.languageeng
dc.publisherAmerican Society for Microbiology
dc.rightsAttribution 4.0 International
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectCell Line, Tumor
dc.subjectHumans
dc.subjectNorwalk virus
dc.subjectIsoxazoles
dc.subjectPyrrolidinones
dc.subjectCysteine Endopeptidases
dc.subjectValine
dc.subjectPhenylalanine
dc.subjectViral Proteins
dc.subjectAntiviral Agents
dc.subjectFluorescence Resonance Energy Transfer
dc.subjectVirus Replication
dc.subjectAmino Acid Sequence
dc.subjectMutation
dc.titleSelection and Characterization of Rupintrivir-Resistant Norwalk Virus Replicon Cells In Vitro.
dc.typeArticle
prism.issueIdentifier5
prism.publicationDate2018
prism.publicationNameAntimicrob Agents Chemother
prism.volume62
dc.identifier.doi10.17863/CAM.23206
dcterms.dateAccepted2018-03-02
rioxxterms.versionofrecord10.1128/AAC.00201-18
rioxxterms.versionVoR
rioxxterms.licenseref.urihttp://creativecommons.org/licenses/by/4.0/
rioxxterms.licenseref.startdate2018-05
dc.contributor.orcidHosmillo, Myra [0000-0002-3514-7681]
dc.identifier.eissn1098-6596
rioxxterms.typeJournal Article/Review
pubs.funder-project-idWellcome Trust (097997/Z/11/Z)
pubs.funder-project-idBiotechnology and Biological Sciences Research Council (BB/N001176/1)
pubs.funder-project-idWellcome Trust (207498/Z/17/Z)
pubs.funder-project-idWellcome Trust (207498/Z/17/Z)
cam.issuedOnline2018-04-26
cam.orpheus.successThu Jan 30 13:00:15 GMT 2020 - The item has an open VoR version.
rioxxterms.freetoread.startdate2100-01-01


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