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Structure-based cleavage mechanism of Thermus thermophilus Argonaute DNA guide strand-mediated DNA target cleavage,PNAS 2014 111 (2) 652-657

发布时间:2014年01月27日

PNAS 2014 111 (2) 652-657; published ahead of print December 27, 2013, doi:10.1073/pnas.1321032111

Structure-based cleavage mechanism of Thermus thermophilus Argonaute DNA guide strand-mediated DNA target cleavage

Gang Sheng, Hongtu Zhao, Jiuyu Wang, Yu Rao, Wenwen Tian, Daan C. Swarts, John van der Oost, Dinshaw J. Patel, and Yanli Wang 

Abstract

We report on crystal structures of ternary Thermus thermophilus Argonaute (TtAgo) complexes with 5′-phosphorylated guide DNA and a series of DNA targets. These ternary complex structures of cleavage-incompatible, cleavage-compatible, and postcleavage states solved at improved resolution up to 2.2 angstrom have provided molecular insights into the orchestrated positioning of catalytic residues, a pair of Mg2+ cations, and the putative water nucleophile positioned for in-line attack on the cleavable phosphate for TtAgo-mediated target cleavage by a RNase H-type mechanism. In addition, these ternary complex structures have provided insights into protein and DNA conformational changes that facilitate transition between cleavage-incompatible and cleavage-compatible states, including the role of a Glu finger in generating a cleavage-competent catalytic Asp-Glu-Asp-Asp tetrad. Following cleavage, the seed segment forms a stable duplex with the complementary segment of the target strand.

相关报道:http://www.ibp.cas.cn/kyjz/zxdt/201401/t20140127_4030213.html

文章链接:http://www.pnas.org/content/111/2.toc

 

 

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