Poliovirus Protein 3CD Is the Active Protease for Processing of the Precursor Protein P1 in vitro

• Published in: Journal of general virology Vol. 69; no. 7; pp. 1627 - 1636
• Main Authors: Jore, Jan, De Geus, Bernard, Jackson, Richard J, Pouwels, Peter H, Enger-Valk, Betty E
• Format: Journal Article
• Language: English
• Published: Reading Soc General Microbiol 01.07.1988; Society for General Microbiology
• Subjects: 3C Viral Proteases; Biological and medical sciences; Capsid - biosynthesis; Capsid Proteins; Cysteine Endopeptidases; DNA - genetics; DNA, Viral - genetics; Endopeptidases - genetics; Endopeptidases - metabolism; Fundamental and applied biological sciences. Psychology; Microbiology; poliovirus; Poliovirus - enzymology; Poliovirus - genetics; Protein Precursors - metabolism; Protein Processing, Post-Translational; Recombinant Fusion Proteins - metabolism; Replicative cycle, interference, host-virus relations, pathogenicity, miscellaneous strains; RNA, Viral - metabolism; Viral Proteins - genetics; Viral Proteins - metabolism; Virology; Virus; Proteins; In vitro transcription; Molecular processing; In vitro translation; Picornaviridae; Proteinase; Enterovirus; Biosynthesis precursor; Poliovirus
• DOI: 10.1099/0022-1317-69-7-1627
• ISSN: 0022-1317

Medical Biological Laboratory TNO, P.O. Box 45, 2280 AA Rijswijk, The Netherlands and 1 Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QW, U.K. A transcription/translation system for generating poliovirus proteins in vitro has been used to assess the proteolytic activity of various polypeptides containing the virus-coded 3C region towards the poliovirus precursor protein P1. Plasmids containing a phage T7 promoter followed by either the complete poliovirus P1 sequence or various sequences containing the 3C region were used for this purpose. We showed that all except one of the 3C-containing polypeptides had a very restricted activity towards P1, generating only a small amount of VP1 and no VP0 or VP3. The only polypeptide capable of fully processing P1 into VP0, VP3 and VP1 in vitro was protein 3CD, consisting of the complete 3C and 3D sequences. Keywords: poliovirus, protein processing, plasmids, T7 Received 16 February 1988; accepted 29 March 1988.