Synthesis and functional activity of tRNAs labeled with fluorescent hydrazides in the D-loop.

RNA
Dongli PanBarry S Cooperman

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.

References

Aug 1, 1979·European Journal of Biochemistry·W Wintermeyer, H G Zachau
Jan 1, 1979·Methods in Enzymology·W WintermeyerH G Zachau
Jul 1, 1974·Proceedings of the National Academy of Sciences of the United States of America·C H Yang, D Söll
Jan 1, 1974·Methods in Enzymology·W Wintermeyer, H G Zachau
Sep 24, 1968·European Journal of Biochemistry·R Thiebe, H G Zachau
May 28, 1967·Journal of Molecular Biology·P Cerutti, N Miller
Apr 3, 2001·Science·M M YusupovH F Noller
Jun 25, 2003·Molecular Cell·Andreas SavelsberghWolfgang Wintermeyer
Aug 20, 2004·Proceedings of the National Academy of Sciences of the United States of America·Scott C BlanchardSteven Chu
Sep 28, 2004·Nature Structural & Molecular Biology·Scott C BlanchardJoseph D Puglisi
Mar 15, 2006·Nature Structural & Molecular Biology·Dongli PanBarry S Cooperman
May 2, 2006·Nature Structural & Molecular Biology·Peter BielingMarina V Rodnina
Jan 16, 2007·Molecular Cell·Ute Kothe, Marina V Rodnina
Feb 24, 2007·Molecular Cell·James B MunroScott C Blanchard
Feb 24, 2007·Molecular Cell·Dongli PanBarry S Cooperman
Jul 27, 2007·RNA·Thu BetteridgeYa-Ming Hou
Aug 19, 2007·Proceedings of the National Academy of Sciences of the United States of America·Tae-Hee LeeSteven Chu
Sep 18, 2007·Journal of Molecular Biology·Christina GrigoriadouBarry S Cooperman
May 2, 2008·The Journal of Biological Chemistry·Dongli PanBarry S Cooperman
May 13, 2008·Molecular Cell·Jingyi FeiRuben L Gonzalez

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Citations

Jun 26, 2009·RNA·Beatriz Llano-SoteloAlexander S Mankin
Sep 21, 2011·Proceedings of the National Academy of Sciences of the United States of America·Hanqing LiuBarry S Cooperman
Oct 5, 2011·Proceedings of the National Academy of Sciences of the United States of America·Chunlai ChenYale E Goldman
Jun 14, 2012·PloS One·Benjamin StevensYale E Goldman
Mar 1, 2013·Protein Engineering, Design & Selection : PEDS·Maxim ChudaevWlodek Mandecki
Mar 17, 2012·Nucleic Acids Research·Gabriel RosenblumBarry S Cooperman
Jun 6, 2013·Bioconjugate Chemistry·Dulce AlonsoBarry S Cooperman
Oct 4, 2012·Chemical Communications : Chem Comm·Takumi IshizukaIchiro Hirao
Nov 26, 2009·FEBS Letters·Rebecca W AlexanderZaida Luthey-Schulten
Aug 16, 2014·ACS Chemical Biology·Wei LiuBarry S Cooperman
Mar 10, 2016·Integrative Biology : Quantitative Biosciences From Nano to Macro·Chengyi TuJanet Zoldan
Aug 30, 2012·Protein Science : a Publication of the Protein Society·Yuhong Wang, Ming Xiao
Mar 29, 2014·Journal of Cellular Physiology·Jiaqi LiuAndrew Leask
Nov 4, 2010·Biophysical Journal·Mediha Esra AltuntopYuhong Wang
May 29, 2012·Biophysical Chemistry·Ming Xiao, Yuhong Wang
May 10, 2011·Molecular Cell·Chunlai ChenBarry S Cooperman
Jun 18, 2016·Proceedings of the National Academy of Sciences of the United States of America·Chunlai ChenYale E Goldman
Jul 5, 2013·Journal of the American Chemical Society·Gabriel RosenblumBarry S Cooperman
Sep 15, 2016·Nucleic Acids Research·Anne PlochowietzAchillefs N Kapanidis
Aug 30, 2017·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Wei LiuBarry S Cooperman
Apr 2, 2013·Nature Structural & Molecular Biology·Chunlai ChenYale E Goldman
May 18, 2018·Nature Chemical Biology·Ivan L VolkovMagnus Johansson
Jun 12, 2020·Nature Communications·Anas KhawajaJoanna Rorbach
Jan 25, 2020·Nucleic Acids Research·Bella KoltunKobi Rosenblum
Jan 9, 2021·Proceedings of the National Academy of Sciences of the United States of America·Martin Y NgBarry S Cooperman
Dec 10, 2020·Drug Discovery Today·Iris AlroyYoni Sheinberger
Apr 4, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Barry S Cooperman

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