Sen1介导非编码RNA转录调控机制的单分子研究

非编码RNA转录终止是基因表达调控领域的基本科学问题，其动态调控是保障基因组结构稳定性和生命体遗传信息有效传递的基础。鉴于该调控过程的复杂性，非编码RNA转录终止的动态过程难以用常规方法解决。本项目利用真核转录研究体系，结合单分子磁镊和单分子荧光技术，研究真核酵母体内的转录终止调控蛋白Sen1的动态过程和分子机制。在单分子水平上，通过研究Sen1在RNA上的行走功能厘清调控前期Sen1搜索和识别RNA聚合酶II（RNAP II）的过程，以及调控后期Sen1和RNAP II相互作用的动态过程，进而阐明真核细胞内非编码RNA转录终止的分子机制。本项目从动态结构的层面探讨蛋白质机器调控转录终止的过程，为人类疾病调控等过程提供定量的科学依据。

Non-coding RNA transcription termination is a key question in gene expression regulation, and maintains gene stability and the efficient transfer of genetic information. Due to its complexity, the kinetics of non-coding RNA transcription termination cannot be well settled via common methods. In this project, we will study the kinetics of Sen1 regulation in yeast by combining our newly developed eukaryotic transcription scaffolds with single-molecule magnetic trapping and fluorescence techniques. From a single-molecule view, we will explore the process of Sen1 searching and targeting RNAP II in the early stage of Sen1 regulation via measuring Sen1 translocation activity on RNA, and the kinetic interactions between Sen1 and RNAP II during the late stage of Sen1 regulation. These results will provide important molecular insights into the non-coding RNA transcription termination in eukaryotes. From a kinetic perspective, this project will underscore the transcription termination processes mediated by molecular machines, and will shed light on quantitative regulation of human diseases.