Investigation of the hepatitis C virus replication complex.
Formation of a membrane-associated replication complex, composed of viral proteins, replicating RNA, altered cellular membranes, and other host factors, is a hallmark of all positive-strand RNA viruses. In the case of HCV, RNA replication takes place in a likely endoplasmic reticulum-derived membrane alteration referred to as the "membranous web." In vitro transcription-translation, membrane extraction and flotation analyses, immunofluorescence microscopy, fluorescent in situ hybridization, and RNA metabolic labeling followed by confocal laser scanning microscopy have yielded insights into the structure and function of the HCV replication complex. We describe these techniques and highlight selected results.
Brass
Volker
V
Department of Medicine II, University of Freiburg, Freiburg, Germany.
Gosert
Rainer
R
Moradpour
Darius
D
D000818:Animals; D001973:Bromodeoxyuridine / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D016174:Hepacivirus / Q000378:metabolism*; D017404:In Situ Hybridization, Fluorescence; D014176:Protein Biosynthesis; D012367:RNA, Viral / Q000096:biosynthesis*; D013194:Staining and Labeling; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000378:metabolism*; D014779:Virus Replication
D012367:RNA, Viral; D014764:Viral Proteins; D001973:Bromodeoxyuridine