Viral DNA and a viral peptide can act as cofactors of adenovirus virion proteinase activity

• Published in: Nature (London) Vol. 361; no. 6409; pp. 274 - 275
• Main Authors: Mangel, Walter F, McGrath, William J, Toledo, Diana L, Anderson, Carl W
• Format: Journal Article
• Language: English
• Published: London Nature Publishing 21.01.1993; Nature Publishing Group
• Subjects: adenovirus; Adenoviruses, Human - enzymology; Amino Acid Sequence; Amino acids; Biological and medical sciences; Coenzymes - isolation & purification; Coenzymes - metabolism; Deoxyribonucleic acid; DNA; DNA, Viral - isolation & purification; DNA, Viral - metabolism; Endopeptidases - isolation & purification; Endopeptidases - metabolism; Fundamental and applied biological sciences. Psychology; Humans; Kinetics; Microbiology; Molecular Sequence Data; Morphology, structure, chemical composition, physicochemical properties; Mutation; Proteins; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Viral Proteins - isolation & purification; Viral Proteins - metabolism; Virion - enzymology; Virology; Viruses; Virus; Human; Adenoviridae; Enzymatic activity; Gene; Enzyme; Proteinase; Mutation; Recombinant protein; Thermal sensibility; Cofactor
• ISSN: 0028-0836; 1476-4687
• DOI: 10.1038/361274a0

Human adenovirus (Ad2), like many other viruses, contains a virion-associated proteinase essential for the synthesis of infectious virus particles. We observed proteinase activity in wild-type virus but not in the ts-1 virus, which contains a mutation in the Ad2 L3 endoprotease gene that confers temperature-sensitive processing of virion precursor proteins. Unexpectedly, we did not observe proteinase activity with purified recombinant endoprotease protein (M(r) 23 K). Purified recombinant endoprotease protein, however, complemented the mutation in ts-1 virions, restoring proteinase activity when mixed together. This implied that cofactors may be required. Here we reconstitute proteinase activity in vitro with three purified viral components: (1) the recombinant endoprotease protein; (2) an 11-amino-acid peptide that originates from the carboxy terminus of pVI, the precursor to virion component VI; and (3) adenovirus DNA. The use of DNA for a proteinase activity is unprecedented.