In vivo and in vitro trans-cleavage activity of hepatitis C virus serine proteinase expressed by recombinant baculoviruses.
By the use of recombinant baculoviruses, the trans-cleavage of hepatitis C virus (HCV) non-structural polyprotein was studied. The viral serine proteinase encoded by the NS3 gene was expressed efficiently in insect cells infected with a baculovirus recombined with HCV cDNA corresponding to amino acids 1046-1243 and the signal sequence of the rabies virus G protein. Coinfection studies showed the in vivo trans-cleavage activity of the expressed protein by the use of a recombinant producing NS5 as a substrate. We also found that the partially purified NS3 serine proteinase prepared from the recombinant-infected cells could cleave NS5A/5B substrate. Characterization of the proteinase obtained wil provide basic knowledge on processing of the HCV polyprotein.
Suzuki
T
T
Department of Virology II, National Institute of Health, Tokyo, Japan.
Sato
M
M
Chieda
S
S
Shoji
I
I
Harada
T
T
Yamakawa
Y
Y
Watabe
S
S
Matsuura
Y
Y
Miyamura
T
T
D000595:Amino Acid Sequence; D000818:Animals; D000956:Antigens, Viral; D001483:Base Sequence; D002460:Cell Line; D015870:Gene Expression; D005814:Genes, Viral; D006023:Glycoproteins / Q000235:genetics; D016174:Hepacivirus / Q000201:enzymology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007313:Insecta; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D011498:Protein Precursors / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals / Q000235:genetics; D011820:Rabies virus / Q000235:genetics; D017421:Sequence Analysis; D012697:Serine Endopeptidases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000235:genetics; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
D000956:Antigens, Viral; D006023:Glycoproteins; D011498:Protein Precursors; D021382:Protein Sorting Signals; D012697:Serine Endopeptidases; D014759:Viral Envelope Proteins; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C064951:glycoprotein G, Rabies virus