title: Efficient aminoacylation of resected RNA helices by class II aspartyl-tRNA synthetase dependent on a single nucleotide.
abstract: We show here that small RNA helices which recapitulate part or all of the acceptor stem of yeast aspartate tRNA are efficiently aminoacylated by cognate class II aspartyl-tRNA synthetase. Aminoacylation is strongly dependent on the presence of the single-stranded G73 'discriminator' identity nucleotide and is essentially insensitive to the sequence of the helical region. Substrates which contain as few as 3 bp fused to G73CCAOH are aspartylated. Their charging is insensitive to the sequence of the loop closing the short helical domains. Aminoacylation of the aspartate mini-helix is not stimulated by a hairpin helix mimicking the anticodon domain and containing the three major anticodon identity nucleotides. A thermodynamic analysis demonstrates that enzyme interactions with G73 in the resected RNA substrates and in the whole tRNA are the same. Thus, if the resected RNA molecules resemble in some way the earliest substrates for aminoacylation with aspartate, then the contemporary tRNA(Asp) has quantitatively retained the influence of the major signal for aminoacylation in these substrates.
authors.affiliation: Unité Structure des Macromolécules Biologiques et Mécanismes de Reconnaissance, Centre National de la Recherche Scientifique, Strasbourg, France.
authors.forename: M
authors.initials: M
authors.lastname: Frugier
authors.forename: C
authors.initials: C
authors.lastname: Florentz
authors.forename: R
authors.initials: R
authors.lastname: Giegé
mesh_terms: D000215:Acylation; D000926:Anticodon; D001226:Aspartate-tRNA Ligase / Q000378:metabolism*; D001224:Aspartic Acid / Q000378:metabolism; D001483:Base Sequence; D005075:Biological Evolution; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012331:RNA, Fungal / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D000926:Anticodon; D012331:RNA, Fungal; D001224:Aspartic Acid; D001226:Aspartate-tRNA Ligase