Skip to content

Research at St Andrews

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

Research output: Contribution to journalArticle

DOI

Open Access permissions

Open

Author(s)

Andrew Woodman, Jamie Arnold, Craig Cameron, David John Evans

School/Research organisations

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.
Close

Details

Original languageEnglish
Pages (from-to)6883-6895
JournalNucleic Acids Research
Volume44
Issue number14
Early online date17 Jun 2016
DOIs
StatePublished - 19 Aug 2016

    Research areas

  • Pollovirus, Picornavirus, Polymerase, Fidelity, Recombination, Evolution

Discover related content
Find related publications, people, projects and more using interactive charts.

View graph of relations

Related by author

  1. CpG and UpA dinucleotides in both coding and non-coding regions of echovirus 7 inhibit replication initiation post-entry

    Fros, J. J., Dietrich, I., Alshaikhahmed, K., Passchier, T. C., Evans, D. J. & Simmonds, P. 29 Sep 2017 In : eLife. 6, 29 p., e29112

    Research output: Contribution to journalArticle

  2. Viruses inhibit CO2 fixation in the most abundant phototrophs on Earth

    Puxty, R. J., Millard, A. D., Evans, D. J. & Scanlan, D. J. 20 Jun 2016 In : Current Biology. 26, 12, p. 1585-1589 6 p.

    Research output: Contribution to journalArticle

  3. The Iflaviruses Sacbrood virus and Deformed wing virus evoke different transcriptional responses in the honeybee which may facilitate their horizontal or vertical transmission

    Ryabov, E. V., Fannon, J. M., Moore, J. D., Wood, G. R. & Evans, D. J. 18 Jan 2016 In : PeerJ. 4, e1591

    Research output: Contribution to journalArticle

  4. Spontaneous deletion of an "ORFanage" region facilitates host adaptation in a "photosynthetic" cyanophage

    Puxty, R. J., Perez-Sepulveda, B., Rihtman, B., Evans, D. J., Millard, A. D. & Scanlan, D. J. 15 Jul 2015 In : PLoS One. 10, 7, 13 p., e0132642

    Research output: Contribution to journalArticle

Related by journal

  1. A type III-B CRISPR–Cas effector complex mediating massive target DNA destruction

    Han, W., Li, Y., Deng, L., Feng, M., Peng, W., Hallstrøm, S., Zhang, J., Peng, N., Liang, Y. X., White, M. F. & She, Q. 28 Feb 2017 In : Nucleic Acids Research. 45, 4, p. 1983-1993 11 p.

    Research output: Contribution to journalArticle

  2. The histone chaperone Vps75 forms multiple oligomeric assemblies capable of mediating exchange between histone H3-H4 tetramers and Asf1-H3-H4 complexes

    Hammond, C. M., Sundaramoorthy, R., Larance, M., Lamond, A., Stevens, M. A., El-Mkami, H., Norman, D. G. & Owen-Hughes, T. 27 Jul 2016 In : Nucleic Acids Research. 44, 13, p. 6157-6172 16 p.

    Research output: Contribution to journalArticle

  3. Mechanism of DNA loading by the DNA repair helicase XPD

    Constantinescu-Aruxandei, D., Petrovic-Stojanovska, B., Penedo, C., White, M. F. & Naismith, J. H. 7 Apr 2016 In : Nucleic Acids Research. 44, 6, p. 2806-2815 10 p.

    Research output: Contribution to journalArticle

  4. Multiple nucleic acid cleavage modes in divergent type III CRISPR systems

    Zhang, J., Graham, S., Tello, A., Liu, H. & White, M. F. 29 Feb 2016 In : Nucleic Acids Research. 44, 4, p. 1789-1799 11 p.

    Research output: Contribution to journalArticle

ID: 243427995