Crucial role of the cGAS N terminus in mediating flowable and functional cGAS–DNA condensate formation via DNA interactions

IF 9.4 1区 综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES
Zhelin Jiang, Fan Shi, Juan Li, Rui Liu, Jinhua Zhou, Zhensheng Zhong, Chaowei Shi, Mingming Ma, ShengQi Xiang, Daxing Gao
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引用次数: 0

Abstract

The DNA-sensing protein cGAS plays a pivotal role in the innate immune response and pathogenesis of various diseases. DNA triggers liquid–liquid phase separation (LLPS) and enhances the enzymatic activity of cGAS. However, the regulatory mechanisms of the disordered N terminus remain unclear. Here, we showed that cGAS Nterm , the N-terminal intrinsic disordered region (IDR) of cGAS, modulates the material properties, specifically the flowability, of the condensed phase of cGAS and is required for full enzymatic activity. Full-length cGAS and cGAS Nterm form liquid droplets in the presence of DNA, while the cGAS catalytic domain forms gel-like solid aggregates with compromised enzymatic activity. Multiple key amino acids responsible for the cGAS Nterm –DNA interaction were identified by NMR spectroscopy as well as other biophysical methods and proven to be critical for the functional LLPS of cGAS both in vitro and in vivo. Interestingly, cGAS Nterm acts in trans to transform the solid aggregates of the cGAS catalytic domain into liquid droplets, subsequently restoring its enzymatic activity. Together, our findings highlight the importance of the IDR of cGAS in LLPS upon DNA stimulation and, more importantly, in modulating the fluidity and permeability of the droplets formed by full-length cGAS, which is crucial for its intact enzymatic activity.
cGAS N端通过DNA相互作用介导可流动和功能性cGAS - DNA凝析物形成的关键作用
dna传感蛋白cGAS在先天免疫反应和多种疾病的发病机制中起着关键作用。DNA触发液-液相分离(LLPS),增强cGAS的酶活性。然而,失调的N端调控机制尚不清楚。在这里,我们发现cGAS的n端本征无序区(IDR) Nterm调节了cGAS凝聚相的材料性质,特别是流动性,并且是充分的酶活性所必需的。全长cGAS和cGAS Nterm在DNA存在下形成液滴,而cGAS催化结构域形成凝胶状固体聚集体,酶活性受损。通过核磁共振波谱和其他生物物理方法鉴定了负责cGAS Nterm -DNA相互作用的多个关键氨基酸,并证明了它们在体外和体内对cGAS的功能LLPS至关重要。有趣的是,cGAS Nterm通过反式作用将cGAS催化结构域的固体聚集体转化为液滴,随后恢复其酶活性。总之,我们的研究结果强调了cGAS在DNA刺激下在LLPS中IDR的重要性,更重要的是,在调节全长cGAS形成的液滴的流动性和渗透性方面,这对其完整的酶活性至关重要。
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来源期刊
CiteScore
19.00
自引率
0.90%
发文量
3575
审稿时长
2.5 months
期刊介绍: The Proceedings of the National Academy of Sciences (PNAS), a peer-reviewed journal of the National Academy of Sciences (NAS), serves as an authoritative source for high-impact, original research across the biological, physical, and social sciences. With a global scope, the journal welcomes submissions from researchers worldwide, making it an inclusive platform for advancing scientific knowledge.
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