期刊
ACS NANO
卷 10, 期 2, 页码 2447-2454出版社
AMER CHEMICAL SOC
DOI: 10.1021/acsnano.5b07257
关键词
Bayesian nanoscopy; RNA polymerase II; cluster analysis; nanostructure dynamics; live cell super-resolution
类别
资金
- National Science Foundation of China [21573013, 21390412, 31271423, 31327901]
- 863 Program [SS2015AA020406]
- CAS
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.
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