title: HCV RNA traffic and association with NS5A in living cells.
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.
authors.affiliation: Department of Molecular and Cellular Biology, Research Centre for Infectious Diseases, University of Adelaide, Adelaide and Centre for Cancer Biology, SA Pathology, Adelaide, SA, Australia.
authors.forename: Guillaume N
authors.initials: GN
authors.lastname: Fiches
authors.affiliation: Department of Molecular and Cellular Biology, Research Centre for Infectious Diseases, University of Adelaide, Adelaide and Centre for Cancer Biology, SA Pathology, Adelaide, SA, Australia.
authors.forename: Nicholas S
authors.initials: NS
authors.lastname: Eyre
authors.affiliation: Department of Molecular and Cellular Biology, Research Centre for Infectious Diseases, University of Adelaide, Adelaide and Centre for Cancer Biology, SA Pathology, Adelaide, SA, Australia.
authors.forename: Amanda L
authors.initials: AL
authors.lastname: Aloia
authors.affiliation: Department of Molecular and Cellular Biology, Research Centre for Infectious Diseases, University of Adelaide, Adelaide and Centre for Cancer Biology, SA Pathology, Adelaide, SA, Australia.
authors.forename: Kylie
authors.initials: K
authors.lastname: Van Der Hoek
authors.affiliation: Systems Immunology, School of Medical Sciences, University of New South Wales, Sydney, NSW, Australia.
authors.forename: Brigit
authors.initials: B
authors.lastname: Betz-Stablein
authors.affiliation: Systems Immunology, School of Medical Sciences, University of New South Wales, Sydney, NSW, Australia.
authors.forename: Fabio
authors.initials: F
authors.lastname: Luciani
authors.affiliation: Institute for Immunology and infectious diseases (IIID), Murdoch University, Perth, WA, Australia.
authors.forename: Abha
authors.initials: A
authors.lastname: Chopra
authors.affiliation: Department of Molecular and Cellular Biology, Research Centre for Infectious Diseases, University of Adelaide, Adelaide and Centre for Cancer Biology, SA Pathology, Adelaide, SA, Australia. Electronic address: michael.beard@adelaide.edu.au.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Beard
mesh_terms: D002460:Cell Line; D003593:Cytoplasm / Q000821:virology; D016174:Hepacivirus / Q000235:genetics / Q000378:metabolism*; D055029:Inverted Repeat Sequences; D012367:RNA, Viral / Q000378:metabolism*; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D014779:Virus Replication; D012324:RNA-Dependent RNA Polymerase
chemical_list: D012367:RNA, Viral; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase
keywords: Hepatitis C Virus; Lipid droplets; Live cell Imaging; NS5A; RNA; Replication; Traffic