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Cas1-Cas2在物理和功能上与DnaK相互作用,以调节CRISPR适应

Tom Killelea¹, Juachi U Dimude², Liu He¹

¹School of Life Sciences, University of Nottingham, UK.

Nucleic acids research

|June 2, 2023

概括

陪伴者DnaK抑制了Cas1-Cas2蛋白质复合体,防止了 prokaryotic 适应性免疫力. 删除DnaK或改变其功能可以恢复免疫力,这表明了防止自我向的机制.

科学领域:

微生物学微生物学
分子生物学分子生物学
遗传学是一个遗传学.

背景情况:

Prokaryotic 适应性免疫依赖 Cas1-Cas2 蛋白质复合体将移动遗传元素 (MGE) DNA 集成到 CRISPR 位点中.
没有预先存在的干扰复合体的新免疫的机制尚未完全理解.

研究的目的:

阐明伴侣DnaK在天真适应和 prokaryotic 适应性免疫的过程中的作用.
研究DnaK如何影响Cas1-Cas2复合体对MGEDNA的向.

主要方法:

使用大肠杆菌 (大肠杆菌) 作为模型系统.
使用基因操纵 (基因删除和突变) 来研究DnaK的功能.
使用显微镜在活细胞中可视化光标记的Cas1.

主要成果:

发现DnaK可以抑制Cas1-Cas2复合体的DNA结合和集成,从而阻碍天真适应.
通过DnaK删除,其基质结合域的突变或菌体羔羊蛋白的表达来逆转天真适应的抑制.
活细胞中Cas1焦点的形成取决于活性DNA复制,在DnaK缺乏的细胞中显著增加.

结论:

DnaK 作为对天真适应的抑制作用,防止 Cas1-Cas2 复合体准宿主染色体.
DnaK的活动对于保持平衡至关重要,只允许在必要时针对MGE.
提出了一个模型,DnaK释放Cas1-Cas2复合体,以针对特定触发器对MGE DNA进行向.

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