title: The discriminator base influences tRNA structure at the end of the acceptor stem and possibly its interaction with proteins.
abstract: For many tRNAs, the discriminator base preceding the CCA sequence at the 3' end is important for aminoacylation. We show that the discriminator base influences the stability of the 1.72 base pair onto which it is stacked. Mutations of the discriminator base from adenosine to cytidine or uridine make the cytidine residue in the C1-G72 base pair of mutant Escherichia coli initiator tRNAs more reactive toward sodium bisulfite, the single-strand-specific reagent. The activity of the enzyme Met-tRNA transformylase toward these and other mutant initiator tRNAs is also consistent with destabilization of the 1.72 base pair in vitro and in vivo. By influencing the strength of the 1.72 base pair, the discriminator base could affect the energetic cost of opening the base pair and modulate the structure of the tRNA near the site of aminoacylation. For some aminoacyl-tRNA synthetases and other proteins that interact with tRNA, these factors could be important for specific recognition and/or formation of the transition state during catalysis.
authors.affiliation: Department of Biology, Massachusetts Institute of Technology, Cambridge 02139-4307.
authors.forename: C P
authors.initials: CP
authors.lastname: Lee
authors.forename: N
authors.initials: N
authors.lastname: Mandal
authors.forename: M R
authors.initials: MR
authors.lastname: Dyson
authors.forename: U L
authors.initials: UL
authors.lastname: RajBhandary
mesh_terms: D000217:Acyltransferases / Q000378:metabolism; D001483:Base Sequence; D006860:Hydrogen Bonding; D019877:Hydroxymethyl and Formyl Transferases; D007700:Kinetics; D008718:Methionine-tRNA Ligase / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D012358:RNA, Transfer, Met / Q000737:chemistry* / Q000378:metabolism; D012261:Ribonucleoproteins / Q000737:chemistry; D013329:Structure-Activity Relationship; D046249:Transfer RNA Aminoacylation
chemical_list: D012358:RNA, Transfer, Met; D012261:Ribonucleoproteins; D019877:Hydroxymethyl and Formyl Transferases; C021561:methionyl-tRNA formyltransferase; D000217:Acyltransferases; D008718:Methionine-tRNA Ligase