Tian Yuanyuan, Jia Qi, Li Meijing, Sia Youyang, Hu Pengjing, Chen Kangjing, Li Ming, Li Xueming, Xu Zigang, Ma Lin, Ye Youpi, Lu Ying, Chen Zhucheng
Department of Dermatology, Beijing Children's Hospital, Capital Medical University, National Center for Children's Health, Beijing 100045, China.
Key Laboratory of Major Diseases in Children, Ministry of Education, Beijing 100045, China.
Life Metab. 2025 Apr 9;4(3):loaf013. doi: 10.1093/lifemeta/loaf013. eCollection 2025 Jun.
Nucleosomes are the fundamental unit of chromatin. Chromatin remodeler plays a crucial role in the regulation of gene expression in eukaryotes. It is involved in important physiological processes, such as development, immune response, and metabolic regulation. During gene expression regulation, chromatin remodelers slide nucleosomes along genomic DNA and play a major role in chromatin organization. Chd1 senses the extranucleosomal linker DNA and controls nucleosome spacing in cells. However, the mechanism of linker DNA sensing by Chd1 is not completely understood. Here, we report the cryo-electron microscope (cryoEM) structures of Chd1 engaging nucleosomes in different states. Chd1 induces two exit-DNA conformations, either fully wrapped or partially unwrapped states. Notably, in the unwrapped conformation, the exit DNA interacts with a positively charged loop of the motor, named the exit-DNA binding loop, and traps Chd1 in the closed state in the ATPase cycle, suggesting attenuation of its remodeling activity. Explored single-molecule fluorescence resonance energy transfer (smFRET) and biochemical data supported the regulation of Chd1 remodeling activity by the exit-DNA conformations, which is important for the linker DNA sensitivity. Mutants of the Chd1 exit-DNA binding loop compromised nucleosome organization in yeast cells. Together, our findings provide valuable insights into Chd1 regulation by exit DNA unwrapping. These results provide a new perspective for the study of cell development and metabolism.
核小体是染色质的基本单位。染色质重塑因子在真核生物基因表达调控中起关键作用。它参与重要的生理过程,如发育、免疫反应和代谢调节。在基因表达调控过程中,染色质重塑因子沿着基因组DNA滑动核小体,并在染色质组织中起主要作用。Chd1感知核小体外的连接DNA并控制细胞中的核小体间距。然而,Chd1感知连接DNA的机制尚未完全了解。在此,我们报告了Chd1与处于不同状态的核小体结合的冷冻电子显微镜(cryoEM)结构。Chd1诱导两种出口DNA构象,即完全包裹或部分解包裹状态。值得注意的是,在解包裹构象中,出口DNA与马达的带正电荷环(称为出口DNA结合环)相互作用,并在ATP酶循环中将Chd1捕获在关闭状态,表明其重塑活性减弱。探索的单分子荧光共振能量转移(smFRET)和生化数据支持出口DNA构象对Chd1重塑活性的调节,这对连接DNA敏感性很重要。Chd1出口DNA结合环的突变体损害了酵母细胞中的核小体组织。总之,我们的发现为通过出口DNA解包裹对ChdI的调节提供了有价值的见解。这些结果为细胞发育和代谢的研究提供了新的视角。