Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Antigens, Viral*
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Base Sequence
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Cell Line
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Gene Expression
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Genes, Viral
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Glycoproteins / genetics
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Hepacivirus / enzymology*
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Humans
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Hydrogen-Ion Concentration
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Insecta
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Molecular Sequence Data
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Nucleopolyhedroviruses / genetics*
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Protein Precursors / metabolism
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Protein Processing, Post-Translational
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Protein Sorting Signals / genetics
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Rabies virus / genetics
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Sequence Analysis
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Serine Endopeptidases / genetics
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Serine Endopeptidases / isolation & purification
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Serine Endopeptidases / metabolism*
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Viral Envelope Proteins / genetics
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Viral Nonstructural Proteins / genetics
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Viral Nonstructural Proteins / isolation & purification
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Viral Nonstructural Proteins / metabolism*
Substances
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Antigens, Viral
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Glycoproteins
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NS3 protein, hepatitis C virus
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Protein Precursors
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Protein Sorting Signals
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Viral Envelope Proteins
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Viral Nonstructural Proteins
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glycoprotein G, Rabies virus
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Serine Endopeptidases