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A three-fluorophore FRET assay for high-throughput screening of small-molecule inhibitors of ribosome assembly
JournalArticle (Originalarbeit in einer wissenschaftlichen Zeitschrift)
 
ID 156758
Author(s) Klostermeier, D.; Sears, P.; Wong, C. -H.; Millar, D. P.; Williamson, J. R.
Author(s) at UniBasel Klostermeier, Dagmar
Year 2004
Title A three-fluorophore FRET assay for high-throughput screening of small-molecule inhibitors of ribosome assembly
Journal Nucleic Acids Research
Volume 32
Number 9
Pages / Article-Number 2707-2715
Keywords Aminoglycosides/chemistry/metabolism; Escherichia coli; Fluorescence; Fluorescence Resonance Energy Transfer/*methods; *Fluorescent Dyes; Magnesium/metabolism; Molecular Structure; Nucleic Acid Conformation; RNA; Bacterial/chemistry/genetics/metabolism; Ribosomal; 16S/chemistry/genetics/metabolism; Ribosomal Proteins/metabolism; Ribosomes/chemistry/genetics/*metabolism
Abstract In one of the first steps of prokaryotic ribosome assembly, the ribosomal protein S15 binds to a three-way junction in the central domain of the 16S rRNA. Binding causes a conformational change that is required for subsequent binding events. Using a novel fluorescence resonance energy transfer assay with three fluorophores, two on the RNA and one on the S15 protein, small-molecule libraries can be screened for potential inhibitors of this initial step in ribosome assembly. The employment of three fluorophores allows both the conformational change of the RNA and the binding of S15 to be monitored in a single assay.
Publisher Oxford University Press
ISSN/ISBN 0305-1048 ; 1362-4962
edoc-URL http://edoc.unibas.ch/dok/A5259713
Full Text on edoc Available
Digital Object Identifier DOI 10.1093/nar/gkh588
ISI-Number WOS:000221746900014
Document type (ISI) Article
 
   

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