Complete replication of a eukaryotic virus RNA in vitro by a purified RNA-dependent RNA polymerase.
A soluble RNA-dependent RNA polymerase was isolated from Nicotiana tabacum plants infected with cucumber mosaic virus (CMV), which has a genome of three positive-strand RNA components, 1, 2, and 3. The purified polymerase contained two virus-encoded polypeptides and one host polypeptide. Polymerase activity was completely dependent on addition of CMV RNA as template, and the products of reaction were single-stranded (ss) RNA and double-stranded (ds) RNA, corresponding to RNAs 1, 2, and 3, and a subgenomic RNA (RNA 4) derived from RNA 3. The ratio of ssRNA to dsRNA was about 5:1, and the ssRNA was shown to be predominantly the positive strand. This demonstrates the complete replication of a eukaryotic virus RNA in vitro by a template-dependent RNA polymerase.
Hayes
R J
RJ
Department of Biology, Imperial College of Science, Technology and Medicine, London, England.
Buck
K W
KW
D001483:Base Sequence; D005814:Genes, Viral; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000201:enzymology / Q000235:genetics*; D016297:Mutagenesis, Site-Directed; D015345:Oligonucleotide Probes; D010947:Plants, Toxic; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D012324:RNA-Dependent RNA Polymerase / Q000302:isolation & purification / Q000378:metabolism*; D014026:Nicotiana
D015345:Oligonucleotide Probes; D012367:RNA, Viral; D012324:RNA-Dependent RNA Polymerase