数字农科院2.0

A mesophilic Argonaute from Clostridium formicaceticum with efficient DNA cleavage activity guided by small DNA

文献类型： 外文期刊

作者： Li, Jianrui;Yu, Meixia;Yang, Zhijia;Zhou, Yueheng;Teng, Yunpeng;Wang, Zijian;Chen, Jian;Lai, Jinsheng;Xin, Beibei

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期刊名称： STRUCTURE

ISSN： 0969-2126

年卷期： 2025 年 33 卷 6 期

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收录情况： SCIE（2025版）

摘要： We characterized a new Argonaute protein (pAgo), CfAgo, from the mesophilic bacterium Clostridium formicaceticum. CfAgo possesses DNA-guided DNA endonuclease activity and cleaves DNA targets at the canonical site. It is active from 28 degrees C to 75 degrees C and prefers DNA guides with a 5'-phosphate group and thymidine as the first nucleotide. Cleavage activity is reduced by single-nucleotide mismatches in the seed, central, and 3'-supplementary regions of guides, with stronger mismatch discrimination observed for 5'hydroxylated (5'OH) guides compared to 5'phosphorylated (5'P) guides. Moreover, structural analysis suggests that the MID domain of CfAgo is crucial for recognizing the 5' guide and it influences the binding specificity. CfAgo catalyzes programmable cleavage of double-stranded DNA in AT-rich regions in the presence of Mn2+ and Mg2+ ions at appropriate salt concentrations. These properties could make CfAgo a promising tool for DNA manipulation such as nucleic acid detection and cleavage.

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