Visualizing carboxyl-terminal domain of RNA polymerase Ⅱ recruitment by FET fusion protein condensates with DNA curtains
Visualizing carboxyl-terminal domain of RNA polymerase Ⅱ recruitment by FET fusion protein condensates with DNA curtains作者机构：Center for Quantitative Biology and Peking-Tsinghua Center for Life Sciences Academy for Advanced Interdiscip-linary Studies Peking University
出 版 物：《Biophysics Reports》 (生物物理学报)
年 卷 期：2022年第8卷第2期
页 面：80-89页
核心收录：
学科分类：0710[理学-生物学] 07[理学] 071007[理学-遗传学]
基 金：supported by the National Natural Science Foundation of China (32088101 and 31670762)
主 题：Liquid–liquid phase separation (LLPS) Biomolecular condensates Single-molecule biophysics DNA curtains In vitro transcription assay
摘 要：Many recent references show that living cells can form some membrane-less organelles by liquid–liquid phase separation(LLPS) of biomolecules, like proteins and nucleic acids. LLPS has been confirmed to link with many important biological functions in living cells, and one of the most important functions of biomolecular condensates is in the field of RNA transcription. Many studies confirm that mammalian RNA polymerase Ⅱ(Pol Ⅱ) molecules containing the CTD with different phosphorylation level are purposed to shuttle between initiation condensates and elongation condensates of RNA transcription. Traditional ensemble assays often experience difficulties in quantitively and directly recording the transient recruitment of Pol Ⅱ CTD. Novel single-molecule approach — DNA curtains can be used to directly visualize biomolecular condensates formation and also recruitment of RNA polymerase Ⅱ(Pol Ⅱ) carboxyl-terminal domain(CTD) at the target sites in solution and in real time. This method can offer the potential for new insights into the mechanism of gene transcription. Here, we highlight the detailed protocol of DNA curtains method for studying LLPS.
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