title: Complex formation between hepatitis C virus NS2 and NS3 proteins.
abstract: Hepatitis C virus (HCV) NS2 and NS3 proteins as well as the NS3 protease cofactor NS4A are essential for the replication of the virus. The presence of in vivo heterodimeric complex between HCV NS2 and NS3 has been suggested by biochemical studies. Detailed characterization of the interactions between these viral proteins is of great importance for better understanding their role in viral replication cycle and represents attractive target for antiviral agents. In this study, we demonstrated in vivo interactions between HCV NS2 and NS3 proteins using an epitope tagging technique. For this purpose NS2, NS3 and NS4A were expressed in fusion with two different tags in Cos7 cells. Immunofluorescence analysis and co-immunoprecipitation with tag-specific antibodies revealed the existence of biologically important NS3/NS4A and NS3/NS2 complexes. Similar complexes were detected also in Huh7 cells infected with Semliki Forest virus vectors expressing NS2 and NS3 or NS23 precursor polyprotein. The formation of complex between NS2 and NS3 was found not to depend on whether the proteins were expressed individually or in form of common precursor. This observation suggests the existence of direct interaction between these two proteins that may have importance for the formation of the whole HCV replication complex.
authors.affiliation: Department of Microbiology and Virology, Institute of Molecular and Cell Biology, Tartu University, Riia Street 23, 51010 Tartu, Estonia. kaja@ebc.ee
authors.forename: Kaja
authors.initials: K
authors.lastname: Kiiver
authors.forename: Andres
authors.initials: A
authors.lastname: Merits
authors.forename: Mart
authors.initials: M
authors.lastname: Ustav
authors.forename: Eva
authors.initials: E
authors.lastname: Zusinaite
mesh_terms: D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D019281:Dimerization; D016174:Hepacivirus / Q000378:metabolism*; D006801:Humans; D047468:Immunoprecipitation; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D013194:Staining and Labeling; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D017361:Viral Nonstructural Proteins; C097010:NS2 protein, Hepatitis C virus; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases