Purification and biochemical characterisation of rabbit calicivirus RNA-dependent RNA polymerases and identification of non-nucleoside inhibitors

Nadezda Urakova, Natalie E Netzler, Andrew G. Kelly, Michael FRESE, Peter A White, Tanja Strive

    Research output: Contribution to journalArticle

    10 Citations (Scopus)

    Abstract

    Rabbit haemorrhagic disease virus (RHDV) is a calicivirus that causes acute infections in both domestic and wild European rabbits (Oryctolagus cuniculus). The virus causes significant economic losses in rabbit farming and reduces wild rabbit populations. The recent emergence of RHDV variants capable of overcoming immunity to other strains emphasises the need to develop universally effective antivirals to enable quick responses during outbreaks until new vaccines become available. The RNA-dependent RNA polymerase (RdRp) is a primary target for the development of such antiviral drugs. In this study, we used cell-free in vitro assays to examine the biochemical characteristics of two rabbit calicivirus RdRps and the effects of several antivirals that were previously identified as human norovirus RdRp inhibitors. The non-nucleoside inhibitor NIC02 was identified as a potential scaffold for further drug development against rabbit caliciviruses. Our experiments revealed an unusually high temperature optimum (between 40 and 45 °C) for RdRps derived from both a pathogenic and a non-pathogenic rabbit calicivirus, possibly demonstrating an adaptation to a host with a physiological body temperature of more than 38 °C. Interestingly, the in vitro polymerase activity of the non-pathogenic calicivirus RdRp was at least two times higher than that of the RdRp of the highly virulent RHDV.
    Original languageEnglish
    Article number100
    Pages (from-to)1-17
    Number of pages17
    JournalViruses
    Volume8
    Issue number4
    DOIs
    Publication statusPublished - 14 Apr 2016

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    RNA Replicase
    Rabbit Haemorrhagic Disease Virus
    Rabbits
    Antiviral Agents
    Norovirus
    Body Temperature
    Agriculture
    Disease Outbreaks
    Immunity
    Vaccines
    Economics
    Viruses
    Temperature
    Infection
    Pharmaceutical Preparations

    Cite this

    Urakova, Nadezda ; Netzler, Natalie E ; Kelly, Andrew G. ; FRESE, Michael ; White, Peter A ; Strive, Tanja. / Purification and biochemical characterisation of rabbit calicivirus RNA-dependent RNA polymerases and identification of non-nucleoside inhibitors. In: Viruses. 2016 ; Vol. 8, No. 4. pp. 1-17.
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    abstract = "Rabbit haemorrhagic disease virus (RHDV) is a calicivirus that causes acute infections in both domestic and wild European rabbits (Oryctolagus cuniculus). The virus causes significant economic losses in rabbit farming and reduces wild rabbit populations. The recent emergence of RHDV variants capable of overcoming immunity to other strains emphasises the need to develop universally effective antivirals to enable quick responses during outbreaks until new vaccines become available. The RNA-dependent RNA polymerase (RdRp) is a primary target for the development of such antiviral drugs. In this study, we used cell-free in vitro assays to examine the biochemical characteristics of two rabbit calicivirus RdRps and the effects of several antivirals that were previously identified as human norovirus RdRp inhibitors. The non-nucleoside inhibitor NIC02 was identified as a potential scaffold for further drug development against rabbit caliciviruses. Our experiments revealed an unusually high temperature optimum (between 40 and 45 °C) for RdRps derived from both a pathogenic and a non-pathogenic rabbit calicivirus, possibly demonstrating an adaptation to a host with a physiological body temperature of more than 38 °C. Interestingly, the in vitro polymerase activity of the non-pathogenic calicivirus RdRp was at least two times higher than that of the RdRp of the highly virulent RHDV.",
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    Purification and biochemical characterisation of rabbit calicivirus RNA-dependent RNA polymerases and identification of non-nucleoside inhibitors. / Urakova, Nadezda; Netzler, Natalie E; Kelly, Andrew G.; FRESE, Michael; White, Peter A; Strive, Tanja.

    In: Viruses, Vol. 8, No. 4, 100, 14.04.2016, p. 1-17.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Purification and biochemical characterisation of rabbit calicivirus RNA-dependent RNA polymerases and identification of non-nucleoside inhibitors

    AU - Urakova, Nadezda

    AU - Netzler, Natalie E

    AU - Kelly, Andrew G.

    AU - FRESE, Michael

    AU - White, Peter A

    AU - Strive, Tanja

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    AB - Rabbit haemorrhagic disease virus (RHDV) is a calicivirus that causes acute infections in both domestic and wild European rabbits (Oryctolagus cuniculus). The virus causes significant economic losses in rabbit farming and reduces wild rabbit populations. The recent emergence of RHDV variants capable of overcoming immunity to other strains emphasises the need to develop universally effective antivirals to enable quick responses during outbreaks until new vaccines become available. The RNA-dependent RNA polymerase (RdRp) is a primary target for the development of such antiviral drugs. In this study, we used cell-free in vitro assays to examine the biochemical characteristics of two rabbit calicivirus RdRps and the effects of several antivirals that were previously identified as human norovirus RdRp inhibitors. The non-nucleoside inhibitor NIC02 was identified as a potential scaffold for further drug development against rabbit caliciviruses. Our experiments revealed an unusually high temperature optimum (between 40 and 45 °C) for RdRps derived from both a pathogenic and a non-pathogenic rabbit calicivirus, possibly demonstrating an adaptation to a host with a physiological body temperature of more than 38 °C. Interestingly, the in vitro polymerase activity of the non-pathogenic calicivirus RdRp was at least two times higher than that of the RdRp of the highly virulent RHDV.

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    KW - Non-nucleoside inhibitors

    KW - Polymerase

    KW - RCV-A1

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    KW - Caliciviridae Infections/drug therapy

    KW - Enzyme Activation/drug effects

    KW - Gene Expression

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    KW - Antiviral Agents/chemistry

    KW - Drug Discovery

    KW - Recombinant Fusion Proteins

    KW - Amino Acid Motifs

    KW - Hemorrhagic Disease Virus, Rabbit/drug effects

    KW - Dose-Response Relationship, Drug

    KW - Animals

    KW - Recombination, Genetic

    KW - Inhibitory Concentration 50

    KW - Kinetics

    KW - Evolution, Molecular

    KW - antiviral agents

    KW - polymerase

    KW - non-nucleoside inhibitors

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