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J. Am. Chem. Soc., 2013, 135 (45), pp 16865–16871 | DOI: 10.1021/ja403138d Dynamic Anchoring of the 3′-End of the Guide Strand Controls the Target Dissociation of Argonaute–Guide Complex

Abstract

Seung-Ryoung Jung^†‡, Eunji Kim^∥, Wonseok Hwang ^†‡, Soochul Shin^§, Ji-Joon Song^*∥, and Sungchul Hohng ^*†‡§

^†Department of Physics and Astronomy, ^‡National Center for Creative Research Initiatives, and ^§Department of Biophysics and Chemical Biology, Seoul National University, Seoul 151-747, Korea
^∥ Department of Biological Sciences, KAIST, Daejeon 305-701 Korea

^*Correspondence to Ji-Joon Song, Sungchul Hohng
S.-R.J. and E.K. contributed equally to this work.

Abstract
Argonaute (Ago) is the catalytic core of small RNA-based gene regulation. Despite plenty of mechanistic studies on Ago, the dynamical aspects and the mechanistic determinants of target mRNA binding and dissociation of Ago-guide strand remain unclear. Here, by using single-molecule fluorescence resonance energy transfer (FRET) assays and Thermus thermophilus Ago (TtAgo), we reveal that the 3′-end of the guide strand dynamically anchors at and releases from the PAZ domain of Ago, and that the 3′-end anchoring of the guide strand greatly accelerates the target dissociation by destabilizing the guide-target duplex. Our results indicate that the target binding/dissociation of Ago?guide is executed through the dynamic interplays among Ago, guide, and target.

논문정보

형식Research article
게재일2013년 11월 (BRIC 등록일 2014-02-07)
연구진국내 연구진
분야 생명과학 > 구조생물 및 생물물리학

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