Effect of conformational features on the aminoacylation of tRNAs and consequences on the permutation of tRNA specificities.
The structure and function of in vitro transcribed tRNA(Asp) variants with inserted conformational features characteristic of yeast tRNA(Phe), such as the length of the variable region or the arrangement of the conserved residues in the D-loop, have been investigated. Although they exhibit significant conformational alterations as revealed by Pb2+ treatment, these variants are still efficiently aspartylated by yeast aspartyl-tRNA synthetase. Thus, this synthetase can accommodate a variety of tRNA conformers. In a second series of variants, the identity determinants of yeast tRNA(Phe) were transplanted into the previous structural variants of tRNA(Asp). The phenylalanine acceptance of these variants improves with increasing the number of structural characteristics of tRNA(Phe), suggesting that phenylalanyl-tRNA synthetase is sensitive to the conformational frame embedding the cognate identity nucleotides. These results contrast with the efficient transplantation of tRNA(Asp) identity elements into yeast tRNA(Phe). This indicates that synthetases respond differently to the detailed conformation of their tRNA substrates. Efficient aminoacylation is not only dependent on the presence of the set of identity nucleotides, but also on a precise conformation of the tRNA.
Perret
V
V
Laboratoire de Biochimie, Institut de Biologie Moléculaire et Cellulaire du CNRS, Strasbourg, France.
Florentz
C
C
Puglisi
J D
JD
Giegé
R
R
D001226:Aspartate-tRNA Ligase / Q000378:metabolism*; D001483:Base Sequence; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010652:Phenylalanine-tRNA Ligase / Q000378:metabolism*; D012331:RNA, Fungal / Q000378:metabolism / Q000648:ultrastructure; D012349:RNA, Transfer, Asp / Q000378:metabolism / Q000648:ultrastructure*; D012360:RNA, Transfer, Phe / Q000378:metabolism / Q000648:ultrastructure*; D012441:Saccharomyces cerevisiae; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D046249:Transfer RNA Aminoacylation
D012331:RNA, Fungal; D012349:RNA, Transfer, Asp; D012360:RNA, Transfer, Phe; D001226:Aspartate-tRNA Ligase; D010652:Phenylalanine-tRNA Ligase