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dc.contributor.authorWoodman, Andrew
dc.contributor.authorArnold, Jamie
dc.contributor.authorCameron, Craig
dc.contributor.authorEvans, David John
dc.date.accessioned2016-06-27T11:30:03Z
dc.date.available2016-06-27T11:30:03Z
dc.date.issued2016-08-19
dc.identifier.citationWoodman , A , Arnold , J , Cameron , C & Evans , D J 2016 , ' Biochemical and genetic analysis of the role of the viral polymerase in enterovirus recombination ' , Nucleic Acids Research , vol. 44 , no. 14 , pp. 6883-6895 . https://doi.org/10.1093/nar/gkw567en
dc.identifier.issn0305-1048
dc.identifier.otherPURE: 243427995
dc.identifier.otherPURE UUID: bb1f527a-6862-4a36-a66e-3ea55763f711
dc.identifier.otherScopus: 84984919845
dc.identifier.otherWOS: 000382999900034
dc.identifier.otherORCID: /0000-0002-1315-4258/work/104252489
dc.identifier.urihttps://hdl.handle.net/10023/9050
dc.descriptionFunding: Biotechnology and Biological Sciences Research Council [BB/M009343/1] and PhD. studentship funding for AW (to D.J.E.); NIH [R01 AI045818 from NIAID to C.E.C.). Funding for open access charge: Biotechnology and Biological Sciences Research Council and University of St. Andrews.en
dc.description.abstractGenetic recombination in single-strand, positive-sense RNA viruses is a poorly understand mechanism responsible for generating extensive genetic change and novel phenotypes. By moving a critical cis-acting replication element (CRE) from the polyprotein coding region to the 3’ non-coding region we have further developed a cell-based assay (the 3’CRE-REP assay) to yield recombinants throughout the non-structural coding region of poliovirus from dually transfected cells. We have additionally developed a defined biochemical assay in which the only protein present is the poliovirus RNA dependent RNA polymerase (RdRp), which recapitulates the strand transfer events of the recombination process. We have used both assays to investigate the role of the polymerase fidelity and nucleotide turnover rates in recombination. Our results, of both poliovirus intertypic and intratypic recombination in the CRE-REP assay and using a range of polymerase variants in the biochemical assay, demonstrate that RdRp fidelity is a fundamental determinant of recombination frequency. High fidelity polymerases exhibit reduced recombination and low fidelity polymerases exhibit increased recombination in both assays. These studies provide the basis for the analysis of poliovirus recombination throughout the non-structural region of the virus genome and provide a defined biochemical assay to further dissect this important evolutionary process.
dc.language.isoeng
dc.relation.ispartofNucleic Acids Researchen
dc.rights© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.en
dc.subjectPollovirusen
dc.subjectPicornavirusen
dc.subjectPolymeraseen
dc.subjectFidelityen
dc.subjectRecombinationen
dc.subjectEvolutionen
dc.subjectQD Chemistryen
dc.subjectQH301 Biologyen
dc.subjectNDASen
dc.subjectBDCen
dc.subjectSDG 3 - Good Health and Well-beingen
dc.subject.lccQDen
dc.subject.lccQH301en
dc.titleBiochemical and genetic analysis of the role of the viral polymerase in enterovirus recombinationen
dc.typeJournal articleen
dc.contributor.sponsorBBSRCen
dc.description.versionPublisher PDFen
dc.contributor.institutionUniversity of St Andrews. School of Biologyen
dc.contributor.institutionUniversity of St Andrews. Biomedical Sciences Research Complexen
dc.identifier.doihttps://doi.org/10.1093/nar/gkw567
dc.description.statusPeer revieweden
dc.identifier.grantnumberBB/M009343/1en


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