Dynamic Anchoring of the 3鈥?End of the Guide Strand Controls the Target Dissociation of Argonaute鈥揋uide Complex

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• 作者：Seung-Ryoung Jung ; Eunji Kim ; Wonseok Hwang ; Soochul Shin ; Ji-Joon Song ; Sungchul Hohng
• 刊名：Journal of the American Chemical Society
• 出版年：2013
• 出版时间：November 13, 2013
• 年：2013
• 卷：135
• 期：45
• 页码：16865-16871
• 全文大小：426K
• 年卷期：v.135,no.45(November 13, 2013)
• ISSN：1520-5126

文摘

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鈥揼uide strand remain unclear. Here, by using single-molecule fluorescence resonance energy transfer (FRET) assays and Thermus thermophilus Ago (TtAgo), we reveal that the 3鈥?end of the guide strand dynamically anchors at and releases from the PAZ domain of Ago, and that the 3鈥?end anchoring of the guide strand greatly accelerates the target dissociation by destabilizing the guide鈥搕arget duplex. Our results indicate that the target binding/dissociation of Ago鈥揼uide is executed through the dynamic interplays among Ago, guide, and target.