HCV RNA traffic and association with NS5A in living cells

Guillaume Fiches, Nicholas Eyre, Amanda Aloia, Kylie Van Der Hoek, Brigit Betz-Stablein, Fabio Luciani, Abha Chopra, Michael Beard

    Research output: Contribution to journalArticlepeer-review

    7 Citations (Scopus)

    Abstract

    The spatiotemporal dynamics of Hepatitis C Virus (HCV) RNA localisation are poorly understood. To address this we engineered HCV genomes harbouring MS2 bacteriophage RNA stem-loops within the 3'-untranslated region to allow tracking of HCV RNA via specific interaction with a MS2-Coat-mCherry fusion protein. Despite the impact of these insertions on viral fitness, live imaging revealed that replication of tagged-HCV genomes induced specific redistribution of the mCherry-tagged-MS2-Coat protein to motile and static foci. Further analysis showed that HCV RNA was associated with NS5A in both static and motile structures while a subset of motile NS5A structures was devoid of HCV RNA. Further investigation of viral RNA traffic with respect to lipid droplets (LDs) revealed HCV RNA-positive structures in close association with LDs. These studies provide new insights into the dynamics of HCV RNA traffic with NS5A and LDs and provide a platform for future investigations of HCV replication and assembly.

    Original languageEnglish
    Pages (from-to)60-74
    Number of pages15
    JournalVirology
    Volume493
    DOIs
    Publication statusPublished - 1 Jun 2016

    Keywords

    • Hepatitis C Virus
    • Lipid droplets
    • Live cell Imaging
    • NS5A
    • Replication
    • RNA
    • Traffic

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