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• W2103353313 abstract "We describe an optimized procedure for replacing the dihydrouridine residues of charged tRNAs with Cy3 and Cy5 dyes linked to a hydrazide group, and demonstrate that the labeled molecules are functional in ribosomal activities including 30S initiation complex formation, EF–Tu-dependent binding to the ribosome, translocation, and polypeptide synthesis. This procedure should be straightforwardly generalizable to the incorporation of other hydrazide-linked fluorophores into tRNA or other dihydrouridine-containing RNAs. In addition, we use a rapid turnover FRET experiment, measuring energy transfer between Cy5-labeled tRNA fMet and Cy3-labeled fMetPhe-tRNA Phe , to obtain direct evidence supporting the hypothesis that the early steps of translocation involve movements of the flexible 3′-single-stranded regions of the tRNAs, with the considerable increase in the distance separating the two tRNA tertiary cores occurring later in the process." @default.
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• W2103353313 date "2008-12-31" @default.
• W2103353313 modified "2023-09-26" @default.
• W2103353313 title "Synthesis and functional activity of tRNAs labeled with fluorescent hydrazides in the D-loop" @default.
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• W2103353313 doi "https://doi.org/10.1261/rna.1257509" @default.
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