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单分子实时3D成像转录周期通过调制干扰计

Guanshi Wang¹, Jesse Hauver², Zachary Thomas³

¹Structural Biology Program, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA; BCMB Graduate Program, Weill Cornell Medicine, Cornell University, New York, NY 10065, USA.

|December 17, 2016

概括

这项研究引入了一种新的超高分辨率成像技术, 在基因转录过程中精确追踪它们的运动和动态, 进步我们对基因组调节的理解.

科学领域:

结构生物学
分子生物物理学
基因组学

背景情况:

基本的细胞过程依赖于复杂的分子机器.
现有的结构生物学工具缺乏研究分子机器动力学的必要分辨率.
了解分子机器运动,形状变化和子单位动力学至关重要.

研究的目的:

开发和展示一种新的3D单分子超分辨率成像技术.
为研究分子机器动力学实现高时空分辨率.
探讨复杂的宏分子机器的动态, 特别是RNA聚合酶.

主要方法:

使用3D单分子超分辨率成像.
使用调制干扰度和相位敏感检测.
实现了<2nm的轴定位精度.

主要成果:

在整个转录周期中可视化了单个Escherichia coli多子单元RNA聚合酶.
剖析了启动-延长过渡的动力学.
在激发器逃逸过程中确定了sigma70 (σ70) 启动因子的命运.

结论:

调制干涉测量使我们能够对分子机器动力学有前所未有的洞察力.
这种技术有助于研究基因组调节中的复杂多分子事务.
这为研究以前具有挑战性的分子过程奠定了基础.

关键词:

RNA聚合酶的使用轴向定位一个全酶干扰测量核孔复合体西格玛系数一个分子超高分辨率成像转录转录周期

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