A Single-Molecule Surface-Based Platform to Detect the Assembly and Function of the Human RNA Polymerase II Transcription Machinery.
A Single-Molecule Surface-Based Platform to Detect the Assembly and Function of the Human RNA Polymerase II Transcription Machinery.
复制标题
一个基于表面的平台，可检测人RNA聚合酶II转录机械的组装和功能。
DOI：
10.1016/j.str.2020.07.009
复制
发表时间：
2020-12-01
期刊：
影响因子：
--
通讯作者：
Pertsinidis A
中科院分区：
文献类型：
--
作者：
Park SR;Hauver J;Zhang Y;Revyakin A;Coleman RA;Tjian R;Chu S;Pertsinidis A
Single-molecule detection and manipulation is a powerful tool for unraveling dynamic biological processes. Unfortunately, success in such experiments is often challenged by tethering the bio-molecule(s) of interest to a biocompatible surface. Here we describe a robust surface passivation method by dense polymer-brush grafting, based on optimized polyethylene-glycol (PEG) deposition conditions, exactly at the lower critical point of an aqueous biphasic PEG-salt system. The increased biocompatibility achieved, compared to PEG deposition in sub-optimal conditions away from the critical point, allowed us to successfully detect the assembly and function of a large macro-molecular machine, a fluorescent-labeled multi-subunit, human RNA Polymerase II Transcription Pre-Initiation Complex, on single, promoter-containing, surface-immobilized DNA molecules. This platform will enable probing the complex biochemistry and dynamics of large, multi-subunit macromolecular assemblies, such as during the initiation of human RNA Pol II transcription, at the single-molecule level. Park et al. develop optimized surface preparation procedures for single-molecule experiments. The increased biocompatibility achieved enables visualizing the assembly and function of a large, multi-component molecular machinery, the human RNA Polymerase II transcription pre-initiation complex, on a natural promoter, at the single-molecule level.
登录
查看更多内容
影响因子：
48
作者：
Hua, Boyang;Han, Kyu Young;Zhou, Ruobo;Kim, Hajin;Shi, Xinghua;Abeysirigunawardena, Sanjaya C.;Jain, Ankur;Singh, Digvijay;Aggarwal, Vasudha;Woodson, Sarah A.;Ha, Taekjip
通讯作者：
Ha, Taekjip
影响因子：
64.5
作者：
Friedman LJ;Gelles J
通讯作者：
Gelles J
DOI：
10.1073/pnas.96.3.893
发表时间：
1999-02-02
影响因子：
11.1
作者：
Ha, TJ;Ting, AY;Weiss, S
通讯作者：
Weiss, S
影响因子：
56.9
作者：
PRIME, KL;WHITESIDES, GM
通讯作者：
WHITESIDES, GM
影响因子：
3.8
作者：
Elting, Mary Williard;Leslie, Sabrina R.;Spudich, James A.
通讯作者：
Spudich, James A.