title: Study of RNA Polymerase II Clustering inside Live-Cell Nuclei Using Bayesian Nanoscopy.
abstract: Nanoscale spatiotemporal clustering of RNA polymerase II (Pol II) plays an important role in transcription regulation. However, dynamics of individual Pol II clusters in live-cell nuclei has not been measured directly, prohibiting in-depth understanding of their working mechanisms. In this work, we studied the dynamics of Pol II clustering using Bayesian nanoscopy in live mammalian cell nuclei. With 50 nm spatial resolution and 4 s temporal resolution, Bayesian nanoscopy allows direct observation of the assembly and disassembly dynamics of individual Pol II clusters. The results not only provide quantifications of Pol II clusters but also shed light on the understanding of cluster formation and regulation. Our study suggests that transcription factories form on-demand and recruit Pol II molecules in their pre-elongation phase. The assembly and disassembly of individual Pol II clusters take place asynchronously. Overall, the methods developed herein are also applicable to studying a wide realm of real-time nanometer-scale nuclear processes in live cells.
authors: State Key Laboratory of Membrane Biology, Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences, Peking University , Beijing 100871, China.|Department of Biomedical Engineering, College of Engineering, Peking University , Beijing 100871, China.
authors: Xuanze
authors: X
authors: Chen
authors: State Key Laboratory of Membrane Biology, Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences, Peking University , Beijing 100871, China.
authors: Mian
authors: M
authors: Wei
authors: State Key Laboratory of Membrane Biology, Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences, Peking University , Beijing 100871, China.|School of Physics, Peking University , Beijing 100871, China.
authors: M Mocarlo
authors: MM
authors: Zheng
authors: Department of Physics, Tsinghua University , Beijing 100084, China.
authors: Jiaxi
authors: J
authors: Zhao
authors: State Key Laboratory of Membrane Biology, Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences, Peking University , Beijing 100871, China.
authors: Huiwen
authors: H
authors: Hao
authors: State Key Laboratory of Membrane Biology, Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences, Peking University , Beijing 100871, China.
authors: Lei
authors: L
authors: Chang
authors: Department of Biomedical Engineering, College of Engineering, Peking University , Beijing 100871, China.
authors: Peng
authors: P
authors: Xi
authors: State Key Laboratory of Membrane Biology, Biodynamic Optical Imaging Center (BIOPIC), School of Life Sciences, Peking University , Beijing 100871, China.
authors: Yujie
authors: Y
authors: Sun
mesh_terms: D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure*; D006801:Humans; D011485:Protein Binding; D012319:RNA Polymerase II / Q000737:chemistry / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D012319:RNA Polymerase II
keywords: Bayesian nanoscopy; RNA polymerase II; cluster analysis; live cell super-resolution; nanostructure dynamics