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Biochemical and genetic analysis of the role of the viral polymerase in enterovirus recombination

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Biochemical and genetic analysis of the role of the viral polymerase in enterovirus recombination. / Woodman, Andrew; Arnold, Jamie; Cameron, Craig; Evans, David John.

In: Nucleic Acids Research, Vol. 44, No. 14, 19.08.2016, p. 6883-6895.

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Woodman, A, Arnold, J, Cameron, C & Evans, DJ 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/gkw567

APA

Woodman, 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, 44(14), 6883-6895. https://doi.org/10.1093/nar/gkw567

Vancouver

Woodman A, Arnold J, Cameron C, Evans DJ. Biochemical and genetic analysis of the role of the viral polymerase in enterovirus recombination. Nucleic Acids Research. 2016 Aug 19;44(14):6883-6895. https://doi.org/10.1093/nar/gkw567

Author

Woodman, Andrew ; Arnold, Jamie ; Cameron, Craig ; Evans, David John. / Biochemical and genetic analysis of the role of the viral polymerase in enterovirus recombination. In: Nucleic Acids Research. 2016 ; Vol. 44, No. 14. pp. 6883-6895.

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@article{bb1f527a68624a36a66e3ea55763f711,
title = "Biochemical and genetic analysis of the role of the viral polymerase in enterovirus recombination",
abstract = "Genetic 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.",
keywords = "Pollovirus, Picornavirus, Polymerase, Fidelity, Recombination, Evolution",
author = "Andrew Woodman and Jamie Arnold and Craig Cameron and Evans, {David John}",
year = "2016",
month = "8",
day = "19",
doi = "10.1093/nar/gkw567",
language = "English",
volume = "44",
pages = "6883--6895",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "OXFORD UNIV PRESS",
number = "14",

}

RIS (suitable for import to EndNote) - Download

TY - JOUR

T1 - Biochemical and genetic analysis of the role of the viral polymerase in enterovirus recombination

AU - Woodman, Andrew

AU - Arnold, Jamie

AU - Cameron, Craig

AU - Evans, David John

PY - 2016/8/19

Y1 - 2016/8/19

N2 - Genetic 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.

AB - Genetic 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.

KW - Pollovirus

KW - Picornavirus

KW - Polymerase

KW - Fidelity

KW - Recombination

KW - Evolution

U2 - 10.1093/nar/gkw567

DO - 10.1093/nar/gkw567

M3 - Article

VL - 44

SP - 6883

EP - 6895

JO - Nucleic Acids Research

T2 - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 14

ER -

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ID: 243427995