Endocytic Rab proteins are required for hepatitis C virus replication complex formation.
During infection, hepatitis C virus (HCV) NS4B protein remodels host membranes to form HCV replication complexes (RC) which appear as foci under fluorescence microscopy (FM). To understand the role of Rab proteins in forming NS4B foci, cells expressing the HCV replicon were examined biochemically and via FM. First, we show that an isolated NS4B-bound subcellular fraction is competent for HCV RNA synthesis. Further, this fraction is differentially enriched in Rab1, 2, 5, 6 and 7. However, when examined via FM, NS4B foci appear to be selectively associated with Rab5 and Rab7 proteins. Additionally, dominant negative (DN) Rab6 expression impairs Rab5 recruitment into NS4B foci. Further, silencing of Rab5 or Rab7 resulted in a significant decrease in HCV genome replication. Finally, expression of DN Rab5 or Rab7 led to a reticular NS4B subcellular distribution, suggesting that endocytic proteins Rab5 and Rab7, but not Rab11, may facilitate NS4B foci formation.
Manna
David
D
Department of Biochemistry and Molecular Biology, The Pennsylvania State University, 308 Althouse Laboratory, University Park, PA 16802, USA.
Aligo
Jason
J
Xu
Chenjia
C
Park
Wei Sun
WS
Koc
Hasan
H
Heo
Won Do
WD
Konan
Kouacou V
KV
D002460:Cell Line; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D016174:Hepacivirus / Q000378:metabolism*; D006801:Humans; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D012324:RNA-Dependent RNA Polymerase / Q000502:physiology; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication / Q000502:physiology*; D020691:rab GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*
D012367:RNA, Viral; D014764:Viral Proteins; D012324:RNA-Dependent RNA Polymerase; D020691:rab GTP-Binding Proteins