Study of RNA Polymerase II Clustering inside Live-Cell Nuclei Using Bayesian Nanoscopy
Study of RNA Polymerase II Clustering inside Live-Cell Nuclei Using Bayesian Nanoscopy
复制标题
DOI：
10.1021/acsnano.5b07257
复制
发表时间：
2016-02-01
期刊：
影响因子：
17.1
通讯作者：
Sun, Yujie
中科院分区：
文献类型：
--
作者：
Chen, Xuanze;Wei, Mian;Sun, Yujie
RNA 聚合酶 II (Pol II) 的纳米级时空聚类在转录调控中发挥着重要作用。然而，活细胞核中单个 Pol II 簇的动态尚未直接测量，阻碍了对其工作机制的深入了解。在这项工作中，我们使用贝叶斯纳米显微镜在活哺乳动物细胞核中研究了 Pol II 聚类的动力学。贝叶斯纳米显微镜具有 50 nm 的空间分辨率和 4 s 的时间分辨率，可以直接观察单个 Pol II 簇的组装和拆卸动态。结果不仅提供了 Pol II 簇的量化，而且还有助于理解簇的形成和调控。我们的研究表明转录工厂按需形成并在预延伸阶段招募 Pol II 分子。各个 Pol II 簇的组装和拆卸是异步进行的。总的来说，本文开发的方法也适用于研究活细胞中实时纳米级核过程的广泛领域。
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.