PMID: 9192624Jun 24, 1997Paper

In vitro and in vivo characterization of novel mRNA motifs that bind special elongation factor SelB

Proceedings of the National Academy of Sciences of the United States of America
S J KlugM Famulok

Abstract

The special elongation factor SelB of Escherichia coli promotes selenocysteine incorporation into formate dehydrogenases. This is thought to be achieved through simultaneous binding to selenocysteyl-tRNASec and, in the case of formate dehydrogenase H, to an fdhF mRNA hairpin structure 3' adjacent to the UGA selenocysteine codon. By in vitro selection, novel RNA sequences ("aptamers"), which can interact tightly and specifically with SelB, were isolated from an RNA library. The library was comprised of mutagenized variants of the wild-type fdhF mRNA hairpin. One-half of the selected sequences contained the apical stem-loop of the fdhF mRNA hairpin highly conserved. Some of the aptamers showed deviations in the primary sequence within this region of the wild-type fdhF hairpin motif while still binding with high affinity to SelB. Binding studies performed with truncated versions of SelB revealed that aptamers binding to different sites on the protein have been selected. To dissect SelB binding to the fdhF hairpin from the overall biological function of this complex, four selected aptamers were analyzed in vivo for UGA readthrough in a lacZ fusion construct. Among these, one promoted UGA readthrough in vivo. Three of the aptamers, ...Continue Reading

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Citations

Sep 25, 2001·BioFactors·M Thanbichler, A Böck
Jan 8, 2005·Chembiochem : a European Journal of Chemical Biology·Michael Famulok, Günter Mayer
Oct 31, 2006·Analytical and Bioanalytical Chemistry·Subash Chandra Bose Gopinath
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Apr 26, 2001·Proceedings of the National Academy of Sciences of the United States of America·G MayerM Famulok
Jul 28, 2004·Proceedings of the National Academy of Sciences of the United States of America·Mirko G TheisMichael Famulok
Mar 31, 1999·Proceedings of the National Academy of Sciences of the United States of America·M BlindM Famulok
Jul 20, 2011·Nature Chemical Biology·Jan L VinkenborgMichael Famulok
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May 1, 2021·Metallomics : Integrated Biometal Science·Michael WellsJohn F Stolz
Nov 3, 2009·Journal of Medicinal Chemistry·Michael Famulok

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