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Type: Journal article
Title: Sensitive luminescent reporter viruses reveal appreciable release of hepatitis C virus NS5A protein into the extracellular environment
Author: Eyre, N.
Aloia, A.
Joyce, M.
Chulanetra, M.
Tyrrell, L.
Beard, M.
Citation: Virology, 2017; 507:20-31
Publisher: Academic Press
Issue Date: 2017
ISSN: 0042-6822
Statement of
Nicholas S. Eyre, Amanda L. Aloia, Michael A. Joyce, Monrat Chulanetra, D. Lorne Tyrrell, Michael R. Bear
Abstract: The HCV NS5A protein is essential for viral RNA replication and virus particle assembly. To study the viral replication cycle and NS5A biology we generated an infectious HCV construct with a NanoLuciferase (NLuc) insertion within NS5A. Surprisingly, beyond its utility as a sensitive reporter of cytoplasmic viral RNA replication, we also observed strong luminescence in cell culture fluids. Further analysis using assembly-defective viruses and subgenomic replicons revealed that infectious virus production was not required for extracellular NS5A-NLuc activity but was associated with enrichment of extracellular NS5A-NLuc in intermediate-density fractions similar to those of exosomes and virus particles. Additionally, BRET analysis indicated that intracellular and extracellular forms of NS5A may adopt differing conformations. Importantly, infection studies using a human liver chimeric mouse model confirmed robust infection in vivo and ready detection of NLuc activity in serum. We hypothesise that the presence of NS5A in extracellular fluids contributes to HCV pathogenesis.
Keywords: Hepatitis C virus; NS5A; Virus replication; Virus assembly; Exosome; Luciferase
Rights: © 2017 Elsevier Inc. All rights reserved.
RMID: 0030067783
DOI: 10.1016/j.virol.2017.04.003
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Appears in Collections:Molecular and Biomedical Science publications

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