Formation of a ligand-assisted complex of two RNA hairpin loops

Changfeng Hong, Takahiro Otabe, Saki Matsumoto, Chikara Dohno, Asako Murata, Masaki Hagihara, Kazuhiko Nakatani

研究成果: ジャーナルへの寄稿学術誌査読

14 被引用数 (Scopus)

抄録

The hairpin structure is one of the most common secondary structures in RNA and holds a central position in the stream of RNA folding from a non-structured RNA to structurally complex and functional ribonucleoproteins. Since the RNA secondary structure is strongly correlated to the function and can be modulated by the binding of small molecules, we have investigated the modulation of RNA folding by a ligand-assisted formation of loop-loop complexes of two RNA hairpin loops. With a ligand (NCT6), designed based on the ligand binding to the G-G mismatches in double-stranded DNA, we successfully demonstrated the formation of both inter- and intra-molecular NCT6-assisted complex of two RNA hairpin loops. NCT6 selectively bound to the two hairpin loops containing (CGG)3 in the loop region. Native polyacrylamide gel electrophoresis analysis of two doubly-labeled RNA hairpin loops clearly showed the formation of intermolecular NCT6-assisted loop-loop complex. Förster resonance energy-transfer studies of RNA constructs containing two hairpin loops, in which each hairpin was labeled with Alexa488 and Cy3 fluorophores, showed the conformational change of the RNA constructs upon binding of NCT6. These experimental data showed that NCT6 simultaneously bound to two hairpin RNAs at the loop region, and can induce the conformational change of the RNA molecule. These data strongly support that NCT6 functions as molecular glue for two hairpin RNAs. Ligand-assisted kissing: With a ligand, NCT6, which binds to the G-G mismatches in double-stranded DNA, inter- and intramolecular NCT6-assisted complexes of two RNA hairpin loops were formed. Native polyacrylamide gel electrophoresis analysis clearly showed the formation of intermolecular NCT6-assisted loop-loop (kissing) complex. FRET studies suggest that NCT6 acts as the molecular glue for two hairpin RNAs (see figure).

本文言語英語
ページ(範囲)5282-5287
ページ数6
ジャーナルChemistry - A European Journal
20
18
DOI
出版ステータス出版済み - 4月 25 2014
外部発表はい

!!!All Science Journal Classification (ASJC) codes

  • 触媒
  • 有機化学

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