title: Proteolytic activity of the plum pox potyvirus NIa-like protein in Escherichia coli.
abstract: The nucleotide sequence of the small nuclear inclusion protein (NIa)-like cistron of plum pox potyvirus (PPV) has been determined. Viral proteolytic activity was expressed in Escherichia coli cells harboring plasmids with a PPV cDNA insert approximately 7000 nt long. Free PPV capsid protein was detected in these cells, but it was not produced when a mutation was introduced in the PPV cDNA insert which induced a Gln to Pro substitution at the large nuclear inclusion protein (NIb)-capsid protein junction. By mutational analysis, the NIa-like protein was determined to be responsible for the proteolytic activity. A Gln to Ser substitution at the presumed NIa-NIb junction, which inhibited proteolytic processing at the carboxyl end of the protease, had no effect on proteolytic cleavage at the NIb-capsid protein junction. In contrast with the high efficiency of proteolytic processing at the NIb-capsid protein cleavage site, processing at the ends of the PPV protease was not complete, suggesting that the PPV polyprotein, like that of other potyviruses, contains cleavage sites with different properties.
authors.affiliation: Centro de Biología Molecular (CSIC-UAM), Universidad Autónoma de Madrid, Spain.
authors.forename: J A
authors.initials: JA
authors.lastname: García
authors.forename: J L
authors.initials: JL
authors.lastname: Riechmann
authors.forename: S
authors.initials: S
authors.lastname: Laín
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D004279:DNA, Viral / Q000235:genetics; D010450:Endopeptidases; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D008969:Molecular Sequence Data; D009154:Mutation; D010447:Peptide Hydrolases / Q000235:genetics / Q000378:metabolism*; D010942:Plant Viruses / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D012328:RNA Viruses / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D004279:DNA, Viral; D014764:Viral Proteins; D010450:Endopeptidases; D010447:Peptide Hydrolases; C059992:nuclear inclusion protein a, mosaic viruses