title: Release of Infectious Hepatitis C Virus from Huh7 Cells Occurs via a trans-Golgi Network-to-Endosome Pathway Independent of Very-Low-Density Lipoprotein Secretion.
abstract: The release of infectious hepatitis C virus (HCV) particles from infected cells remains poorly characterized. We previously demonstrated that virus release is dependent on the endosomal sorting complex required for transport (ESCRT). Here, we show a critical role of trans-Golgi network (TGN)-endosome trafficking during the assembly, but principally the secretion, of infectious virus. This was demonstrated by both small interfering RNA (siRNA)-mediated silencing of TGN-associated adaptor proteins and a panel of dominant negative (DN) Rab GTPases involved in TGN-endosome trafficking steps. Importantly, interfering with factors critical for HCV release did not have a concomitant effect on secretion of triglycerides, ApoB, or ApoE, indicating that particles are likely released from Huh7 cells via pathways distinct from that of very-low-density lipoprotein (VLDL). Finally, we show that HCV NS2 perturbs TGN architecture, redistributing TGN membranes to closely associate with HCV core protein residing on lipid droplets. These findings support the notion that HCV hijacks TGN-endosome trafficking to facilitate particle assembly and release. Moreover, although essential for assembly and infectivity, the trafficking of mature virions is seemingly independent of host lipoproteins.
  
  
  
  The mechanisms by which infectious hepatitis C virus particles are assembled and released from the cell are poorly understood. We show that the virus subverts host cell trafficking pathways to effect the release of virus particles and disrupts the structure of the Golgi apparatus, a key cellular organelle involved in secretion. In addition, we demonstrate that the mechanisms used by the virus to exit the cell are distinct from those used by the cell to release lipoproteins, suggesting that the virus effects a unique modification to cellular trafficking pathways.
authors.affiliation: School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds, United Kingdom.
authors.forename: Jamel
authors.initials: J
authors.lastname: Mankouri
authors.affiliation: School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds, United Kingdom.
authors.forename: Cheryl
authors.initials: C
authors.lastname: Walter
authors.affiliation: School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds, United Kingdom.
authors.forename: Hazel
authors.initials: H
authors.lastname: Stewart
authors.affiliation: Leeds Institute of Cancer and Pathology, Faculty of Medicine and Health, St. James' University Hospital, Beckett St., Leeds, United Kingdom.
authors.forename: Matthew
authors.initials: M
authors.lastname: Bentham
authors.affiliation: Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea.
authors.forename: Wei Sun
authors.initials: WS
authors.lastname: Park
authors.affiliation: Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea.
authors.forename: Won Do
authors.initials: WD
authors.lastname: Heo
authors.affiliation: Graduate School of Life Sciences, Tohoku University, Sendai, Miyagi, Japan.
authors.forename: Mitsunori
authors.initials: M
authors.lastname: Fukuda
authors.affiliation: Leeds Institute of Cancer and Pathology, Faculty of Medicine and Health, St. James' University Hospital, Beckett St., Leeds, United Kingdom s.d.c.griffin@leeds.ac.uk m.harris@leeds.ac.uk.
authors.forename: Stephen
authors.initials: S
authors.lastname: Griffin
authors.affiliation: School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds, United Kingdom s.d.c.griffin@leeds.ac.uk m.harris@leeds.ac.uk.
authors.forename: Mark
authors.initials: M
authors.lastname: Harris
mesh_terms: D001692:Biological Transport; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D011992:Endosomes / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D006056:Golgi Apparatus / Q000235:genetics / Q000378:metabolism / Q000821:virology; D016174:Hepacivirus / Q000502:physiology; D006526:Hepatitis C / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D006801:Humans; D008079:Lipoproteins, VLDL / Q000378:metabolism*; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D008856:Microscopy, Fluorescence; D022142:Secretory Vesicles / Q000378:metabolism; D014771:Virion / Q000378:metabolism; D057074:Virus Release / Q000502:physiology*; D014779:Virus Replication; D021601:trans-Golgi Network / Q000235:genetics / Q000378:metabolism* / Q000821:virology
chemical_list: D008079:Lipoproteins, VLDL