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

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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

PY - 2016/4/14

Y1 - 2016/4/14

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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.

KW - Antiviral agents

KW - Non-nucleoside inhibitors

KW - Polymerase

KW - RCV-A1

KW - RHDV

KW - Amino Acid Sequence

KW - Caliciviridae Infections/drug therapy

KW - Enzyme Activation/drug effects

KW - Gene Expression

KW - RNA Replicase/antagonists & inhibitors

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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U2 - 10.3390/v8040100

DO - 10.3390/v8040100

M3 - Article

VL - 8

SP - 1

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JO - Viruses

JF - Viruses

SN - 1999-4915

IS - 4

M1 - 100

ER -