Identification of the domain required for trans-cleavage activity of hepatitis C viral serine proteinase.
A serine proteinase, Cpro-2, encoded in the hepatitis C virus (HCV) genome, is considered to be located in the N-terminal part of HCV p70, one of the putative nonstructural (NS) proteins of HCV. Cpro-2 is suggested to be responsible for producing several kinds of NS proteins by processing of the HCV precursor polyprotein. We identified the active domain of Cpro-2 and clarified the mechanism of HCV polyprotein processing; various HCV mutants deleted around this serine proteinase structure were cosynthesized with unprocessed HCV polypeptides containing Cpro-2-dependent cleavage sites in COS-1 cells. We showed that Cpro-2 cleaved the HCV precursor polyprotein intermolecularly (trans) and that Cpro-2 domain which is necessary and sufficient for that cleavage mapped to within 167 aa, from Gly1049 to Ser1215 of the HCV precursor polyprotein.
Tanji
Y
Y
Virology Division, National Cancer Center Research Institute, Tokyo, Japan.
Hijikata
M
M
Hirowatari
Y
Y
Shimotohno
K
K
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D004252:DNA Mutational Analysis; D010450:Endopeptidases / Q000235:genetics* / Q000378:metabolism*; D016174:Hepacivirus / Q000201:enzymology*; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000378:metabolism; D017384:Sequence Deletion; D013379:Substrate Specificity; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis
D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins; D010450:Endopeptidases; C084816:proteinase Cpro-2, hepatitis C virus