Publications
2002
Walter F, Putz J, Giege R, Westhof E
Binding of tobramycin leads to conformational changes in yeast tRNA(Asp) and inhibition of aminoacylation Article de journal
Dans: EMBO J, vol. 21, no. 4, p. 760-768, 2002, ISBN: 11847123, (0261-4189 Journal Article).
Résumé | Liens | BibTeX | Étiquettes: Acylation Base Sequence Carbohydrate Sequence Fluorescence Polarization Molecular Sequence Data *Nucleic Acid Conformation RNA, Asp/chemistry/*metabolism Saccharomyces cerevisiae/*genetics Support, Fungal/chemistry/*metabolism RNA, Non-U.S. Gov't Tobramycin/*metabolism, Transfer, Unité ARN, WESTHOF
@article{,
title = {Binding of tobramycin leads to conformational changes in yeast tRNA(Asp) and inhibition of aminoacylation},
author = {F Walter and J Putz and R Giege and E Westhof},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11847123},
isbn = {11847123},
year = {2002},
date = {2002-01-01},
journal = {EMBO J},
volume = {21},
number = {4},
pages = {760-768},
abstract = {Aminoglycosides inhibit translation in bacteria by binding to the A site in the ribosome. Here, it is shown that, in yeast, aminoglycosides can also interfere with other processes of translation in vitro. Steady-state aminoacylation kinetics of unmodified yeast tRNA(Asp) transcript indicate that the complex between tRNA(Asp) and tobramycin is a competitive inhibitor of the aspartylation reaction with an inhibition constant (K(I)) of 36 nM. Addition of an excess of heterologous tRNAs did not reverse the charging of tRNA(Asp), indicating a specific inhibition of the aspartylation reaction. Although magnesium ions compete with the inhibitory effect, the formation of the aspartate adenylate in the ATP-PP(i) exchange reaction by aspartyl-tRNA synthetase in the absence of the tRNA is not inhibited. Ultraviolet absorbance melting experiments indicate that tobramycin interacts with and destabilizes the native L-shaped tertiary structure of tRNA(Asp). Fluorescence anisotropy using fluorescein-labelled tobramycin reveals a stoichiometry of one molecule bound to tRNA(Asp) with a K(D) of 267 nM. The results indicate that aminoglycosides are biologically effective when their binding induces a shift in a conformational equilibrium of the RNA.},
note = {0261-4189
Journal Article},
keywords = {Acylation Base Sequence Carbohydrate Sequence Fluorescence Polarization Molecular Sequence Data *Nucleic Acid Conformation RNA, Asp/chemistry/*metabolism Saccharomyces cerevisiae/*genetics Support, Fungal/chemistry/*metabolism RNA, Non-U.S. Gov't Tobramycin/*metabolism, Transfer, Unité ARN, WESTHOF},
pubstate = {published},
tppubtype = {article}
}
Aminoglycosides inhibit translation in bacteria by binding to the A site in the ribosome. Here, it is shown that, in yeast, aminoglycosides can also interfere with other processes of translation in vitro. Steady-state aminoacylation kinetics of unmodified yeast tRNA(Asp) transcript indicate that the complex between tRNA(Asp) and tobramycin is a competitive inhibitor of the aspartylation reaction with an inhibition constant (K(I)) of 36 nM. Addition of an excess of heterologous tRNAs did not reverse the charging of tRNA(Asp), indicating a specific inhibition of the aspartylation reaction. Although magnesium ions compete with the inhibitory effect, the formation of the aspartate adenylate in the ATP-PP(i) exchange reaction by aspartyl-tRNA synthetase in the absence of the tRNA is not inhibited. Ultraviolet absorbance melting experiments indicate that tobramycin interacts with and destabilizes the native L-shaped tertiary structure of tRNA(Asp). Fluorescence anisotropy using fluorescein-labelled tobramycin reveals a stoichiometry of one molecule bound to tRNA(Asp) with a K(D) of 267 nM. The results indicate that aminoglycosides are biologically effective when their binding induces a shift in a conformational equilibrium of the RNA.