Identification of the hepatitis C virus RNA replication complex in Huh-7 cells harboring subgenomic replicons.
Formation of a membrane-associated replication complex, composed of viral proteins, replicating RNA, and altered cellular membranes, is a characteristic feature of plus-strand RNA viruses. Here, we demonstrate the presence of a specific membrane alteration, designated the membranous web, that contains hepatitis C virus (HCV) nonstructural proteins, as well as viral plus-strand RNA, in Huh-7 cells harboring autonomously replicating subgenomic HCV RNAs. Metabolic labeling with 5-bromouridine 5'-triphosphate in the presence of actinomycin D revealed that the membranous web is the site of viral RNA synthesis and therefore represents the replication complex of HCV.
Gosert
Rainer
R
Department of Medicine II, University of Freiburg, Germany.
Egger
Denise
D
Lohmann
Volker
V
Bartenschlager
Ralf
R
Blum
Hubert E
HE
Bienz
Kurt
K
Moradpour
Darius
D
D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure / Q000821:virology; D016174:Hepacivirus / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D012367:RNA, Viral / Q000096:biosynthesis* / Q000235:genetics / Q000648:ultrastructure*; D012093:Replicon; D014407:Tumor Cells, Cultured; D017361:Viral Nonstructural Proteins / Q000378:metabolism; D014779:Virus Replication; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; D012324:RNA-Dependent RNA Polymerase
D012367:RNA, Viral; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase