1. HCV RNA traffic and association with NS5A in living cells.
- Author
-
Fiches, Guillaume N., Eyre, Nicholas S., Aloia, Amanda L., Van Der Hoek, Kylie, Betz-Stablein, Brigit, Luciani, Fabio, Chopra, Abha, and Beard, Michael R.
- Subjects
- *
HEPATITIS C virus , *BACTERIOPHAGES , *VIRAL genomes , *SPATIOTEMPORAL processes , *RNA , *COAT proteins (Viruses) , *VIRAL replication - 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. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF