Autoproteolytic cleavage of recombinant 3C proteinase of hepatitis A virus.
A hepatitis A virus cDNA fragment coding for the viral proteinase 3C was expressed as a chimeric protein fused in-frame to the C-terminus of beta-galactosidase. Following induction of the lac Z promoter, polypeptides of 150, 28, 26, and 16 kDa, all of which carry 3C antigenicity, were produced. The 28- and 26-kDa proteins were identified as autoproteolytic products of the fusion protein by determination of their N-terminal amino acid sequence. The 16-kDa protein arises from internal initiation. Following substitution of the 37 amino acids at the C-terminus of 3C, the autolytic activity was no longer observed. The recombinant proteinase did not show trans-activity when recombinant proteins of the P1 or P2 region were used as substrates. Antisera directed against recombinant 3C could not detect 3C or its precursors in HAV-infected cells.
Gauss-Müller
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V
Institut für Medizinische Mikrobiologie, Medizinische Universität zu Lübeck, Germany.
Jürgensen
D
D
Deutzmann
R
R
D000086042:3C Viral Proteases; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D006507:Hepatovirus / Q000201:enzymology* / Q000378:metabolism; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000378:metabolism; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism; D015183:Restriction Mapping; D014764:Viral Proteins
D010455:Peptides; D011506:Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins; D003546:Cysteine Endopeptidases; D000086042:3C Viral Proteases