The prokaryotic Argonaute proteins enhance homology sequence-directed recombination in bacteria
The prokaryotic Argonaute proteins enhance homology sequence-directed recombination in bacteria
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原核 Argonaute 蛋白增强细菌中的同源序列定向重组。
DOI：
10.1093/nar/gkz040
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发表时间：
2019-04-23
影响因子：
14.9
通讯作者：
Zhang, Anding
中科院分区：
文献类型：
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作者：
Fu, Lei;Xie, Caiyun;Zhang, Anding
Argonaute 蛋白存在于生命的各个领域并保守存在。最近鉴定的原核 Argonaute 蛋白 (pAgos) 通过 DNA 干扰参与宿主防御。在这里，我们报告说，Natronobacter gregoryi Argonaute (NgAgo) 能够以 80-100% 的效率增强多杀性巴斯德氏菌和大肠杆菌中的基因插入或缺失。此外，这些效应以同源臂依赖性方式存在，但与引导 DNA 和潜在酶活性无关。有趣的是，在其他 pAgos 片段中也观察到了这种效应，包括嗜热栖热菌 Argonaute (TtAgo)、Aquifex aeolicus Argonaute (AaAgo) 和激烈火球菌 Argonaute (PfAgo)。 NgAgo 系统的基本机制是正选择过程，主要通过其 PIWI 样结构域与重组酶 A (recA) 相互作用来增强 recA 介导的 DNA 链交换。我们的研究揭示了一种增强细菌同源序列引导基因编辑的新系统。
Argonaute proteins are present and conserved in all domains of life. Recently characterized prokaryotic Argonaute proteins (pAgos) participates in host defense by DNA interference. Here, we report that the Natronobacterium gregoryi Argonaute (NgAgo) enhances gene insertions or deletions in Pasteurella multocida and Escherichia coli at efficiencies of 80-100%. Additionally, the effects are in a homologous arms-dependent but guide DNA- and potential enzyme activity-independent manner. Interestingly, such effects were also observed in other pAgos fragments including Thermus thermophilus Argonaute (TtAgo), Aquifex aeolicus Argonaute (AaAgo) and Pyrococcus furiosus Argonaute (PfAgo). The underlying mechanism of the NgAgo system is a positive selection process mainly through its PIWI-like domain interacting with recombinase A (recA) to enhance recA-mediated DNA strand exchange. Our study reveals a novel system for enhancing homologous sequence-guided gene editing in bacteria.