title: Determinants in tRNA for activation of arginyl-tRNA synthetase: evidence that tRNA flexibility is required for the induced-fit mechanism.
abstract: Arginyl-tRNA synthetase (ArgRS) catalyzes formation of arginyl-adenylate in a tRNA-dependent reaction. Previous studies have revealed that conformational changes occur upon tRNA binding. In this study, we analyzed the sequence and structural features of tRNA that are essential to activate the catalytic center of mammalian arginyl-tRNA synthetase. Here, tRNA variants with different activator potential are presented. The three regions that are crucial for activation of ArgRS are the terminal adenosine, the D-loop, and the anticodon stem-loop of tRNA. The Add-1 N-terminal domain of ArgRS, which has the very unique property among aminoacyl-tRNA synthetases to interact with the D-loop in the corner of the convex side of tRNA, has an essential role in anchoring tRNA and participating in tRNA-induced amino acid activation. The results suggest that locking the acceptor extremity, the anticodon loop, and the D-loop of tRNA on the catalytic, anticodon-binding, and Add-1 domains of ArgRS also requires some flexibility of the tRNA molecule, provided by G:U base pairs, to achieve the productive conformation of the active site of the enzyme by induced fit.
authors.affiliation: Laboratoire d'Enzymologie et Biochimie Structurales, CNRS, 1 Avenue de la Terrasse, 91190 Gif-sur-Yvette, France.
authors.forename: Ludovic
authors.initials: L
authors.lastname: Guigou
authors.forename: Marc
authors.initials: M
authors.lastname: Mirande
mesh_terms: D001126:Arginine-tRNA Ligase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D002240:Carbohydrate Sequence; D020134:Catalytic Domain; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007093:Imidazoles / Q000494:pharmacology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012343:RNA, Transfer / Q000737:chemistry*
chemical_list: D004791:Enzyme Inhibitors; D007093:Imidazoles; D012343:RNA, Transfer; D001126:Arginine-tRNA Ligase