title: Mutagenesis and self-ligation of the self-cleavage domain of the satellite RNA minus strand of tobacco ringspot virus and its binding to polyamines.
abstract: Several mutants for the minus strands of the self-cleaving domain of the satellite RNA of tobacco ringspot virus have been synthesized by joining chemically synthesized oligoribonucleotides with RNA ligase. Kinetic properties of the enzyme strands (50 nucleotides) against substrates (15-mer and 18-mer) were investigated. Structural properties of the unpaired part in the cleavage region were estimated from mutagenesis. The catalytic domain alone was proved to be responsible for the rejoining reaction of cleaved substrates. It was also found that the ribozyme could be divided into two strands without loss of activity. Effects of concentration of magnesium ion and polyamines on the cleavage reaction for the two-stranded ribozyme are also reported.
authors.affiliation: Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan.
authors.forename: A
authors.initials: A
authors.lastname: Sekiguchi
authors.forename: Y
authors.initials: Y
authors.lastname: Komatsu
authors.forename: M
authors.initials: M
authors.lastname: Koizumi
authors.forename: E
authors.initials: E
authors.lastname: Ohtsuka
mesh_terms: D001483:Base Sequence; D004722:Endoribonucleases / Q000378:metabolism; D007700:Kinetics; D008274:Magnesium / Q000378:metabolism; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides / Q000235:genetics / Q000378:metabolism; D010942:Plant Viruses / Q000235:genetics*; D011073:Polyamines / Q000378:metabolism; D012313:RNA / Q000378:metabolism*; D016337:RNA, Catalytic / Q000378:metabolism*; D019244:RNA, Satellite; D013696:Temperature
chemical_list: D009838:Oligodeoxyribonucleotides; D011073:Polyamines; D016337:RNA, Catalytic; D019244:RNA, Satellite; D012313:RNA; D004722:Endoribonucleases; C022544:ribonuclease U2; D008274:Magnesium