title: In vitro activity of the hairpin ribozyme derived from the negative strand of arabis mosaic virus satellite RNA.
abstract: The negative strand of the satellite RNA of tobacco ringspot virus [(-)sTRSV] is a self-cleaving RNA, of which self-cleaving domain is called the hairpin ribozyme. The negative strand of the satellite RNA of arabis mosaic virus [(-)sArMV] has been suggested to have a hairpin ribozyme-like secondary structure, and we have previously shown that this hairpin domain of (-)sArMV has ribozyme activity. Here we report characterization of the cleavage reaction of the (-)sArMV hairpin ribozyme. Mutagenesis analyses in a trans-acting system revealed, surprisingly, that the wild-type ribozyme was less active than almost all the other mutant ribozymes tested. In a cis-acting system (self-cleaving reaction), however, the reaction of the RNA containing the wild-type sequence proceeds highly efficiently. This result suggests that the inefficient cleavage of the wild-type substrate in trans-acting system may be due to low efficiency at the substrate-binding step but not at the chemical cleavage step in the reaction. We also constructed a chimeric ribozyme between the catalytic hairpin domain from (-)sArMV and the substrate-binding site from (-)sTRSV. This chimeric ribozyme had the highest activity among the trans-acting hairpin ribozymes tested.
authors.affiliation: College of Environmental Health, Azabu University, Sagamihara, Kanagawa.
authors.forename: S
authors.initials: S
authors.lastname: Hisamatsu
authors.forename: Y
authors.initials: Y
authors.lastname: Morikawa
authors.forename: R
authors.initials: R
authors.lastname: Tomita
authors.forename: T
authors.initials: T
authors.lastname: Tanaka
authors.forename: S
authors.initials: S
authors.lastname: Sonoki
authors.forename: Y
authors.initials: Y
authors.lastname: Kikuchi
mesh_terms: D001483:Base Sequence; D007700:Kinetics; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000201:enzymology / Q000235:genetics*; D017860:Nepovirus / Q000737:chemistry; D009690:Nucleic Acid Conformation; D016337:RNA, Catalytic / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019244:RNA, Satellite / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins
chemical_list: D016337:RNA, Catalytic; D019244:RNA, Satellite; D011993:Recombinant Fusion Proteins