title: Nontemplated terminal nucleotidyltransferase activity of double-stranded RNA bacteriophage phi6 RNA-dependent RNA polymerase.
abstract: The replication and transcription of double-stranded RNA (dsRNA) viruses occur within a polymerase complex particle in which the viral genome is enclosed throughout the entire life cycle of the virus. A single protein subunit in the polymerase complex is responsible for the template-dependent RNA polymerization activity. The isolated polymerase subunit of the dsRNA bacteriophage phi6 was previously shown to replicate and transcribe given RNA molecules. In this study, we show that this enzyme also catalyzes nontemplated nucleotide additions to single-stranded and double-stranded nucleic acid molecules. This terminal nucleotidyltransferase activity not only is a property of the isolated enzyme but also is detected to take place within the viral nucleocapsid. This is the first time terminal nucleotidyltransferase activity has been reported for a dsRNA virus as well as for a viral particle. The results obtained together with previous high-resolution structural data on the phi6 RNA-dependent RNA polymerase suggest a mechanism for terminal nucleotidyl addition. We propose that the activity is involved in the termination of the template-dependent RNA polymerization reaction on the linear phi6 genome.
authors.affiliation: Institute of Biotechnology and Department of Biological and Environmental Sciences, P.O. Box 56 (Viikinkaari 5), 00014 University of Helsinki, Finland.
authors.forename: Minna M
authors.initials: MM
authors.lastname: Poranen
authors.forename: Minni R L
authors.initials: MR
authors.lastname: Koivunen
authors.forename: Dennis H
authors.initials: DH
authors.lastname: Bamford
mesh_terms: D017126:Bacteriophage phi 6 / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D008954:Models, Biological; D008969:Molecular Sequence Data; D019251:Nucleocapsid / Q000378:metabolism; D009711:Nucleotides / Q000378:metabolism*; D009713:Nucleotidyltransferases / Q000378:metabolism*; D010957:Plasmids; D012330:RNA, Double-Stranded / Q000235:genetics / Q000378:metabolism*; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics / Q000378:metabolism*; D013698:Templates, Genetic
chemical_list: D009711:Nucleotides; D012330:RNA, Double-Stranded; D012367:RNA, Viral; D009713:Nucleotidyltransferases; D012324:RNA-Dependent RNA Polymerase