title: A single metal ion plays structural and chemical roles in an aminoacyl-transferase ribozyme.
abstract: Catalytically active RNA molecules rely on metal ions for structural and/or catalytic functions. Our in vitro selected aminoacyl-transferase ribozyme is no exception, as it employs a single fully hydrated Mg2+ ion for catalysis [Suga, H., et al. (1998) Biochemistry 37, 10118-10125]. Here we report the essential catalytic residues of the ribozyme and their spatial arrangement in the relation to the metal binding site. Evidence obtained using a combination of Pb2+ and Tb3+ hydrolytic cleavage assays on wild type and mutant ribozymes revealed a cooperative metal binding site that consists of the tandem G:U wobble pairs in P1 and consecutive G:U and U:A pairs in P3. The formation of this concerted Mg2+ binding site positions the P1 and P3 helices in a parallel manner, placing the L3 tetraloop in close proximity to the internal guide sequence (IGS, substrate binding site), which is adjacent to P1. Certain monovalent metal ions inhibit catalysis at low concentrations but support catalysis at high concentrations. These analyses imply that the Mg2+ ion plays both structural and chemical roles and that it brings about the significant rate acceleration in aminoacyl-transfer in concert with the L3-IGS long-range interaction.
authors.affiliation: Department of Chemistry, 657 Natural Sciences Complex, University at Buffalo, State University of New York, Buffalo, New York 14260-3000, USA.
authors.forename: A
authors.initials: A
authors.lastname: Flynn-Charlebois
authors.forename: N
authors.initials: N
authors.lastname: Lee
authors.forename: H
authors.initials: H
authors.lastname: Suga
mesh_terms: D019881:Aminoacyltransferases / Q000737:chemistry* / Q000235:genetics; D001483:Base Sequence; D017124:Conserved Sequence; D007854:Lead / Q000737:chemistry; D008274:Magnesium / Q000737:chemistry*; D009154:Mutation; D009690:Nucleic Acid Conformation; D016337:RNA, Catalytic / Q000737:chemistry* / Q000235:genetics; D013725:Terbium / Q000737:chemistry
chemical_list: D016337:RNA, Catalytic; D013725:Terbium; D007854:Lead; D019881:Aminoacyltransferases; D008274:Magnesium