Self-organization, molecular mechanics, and catalytic functions of nucleoprotein complexes studied using single-DNA micromanipulation

本项目致力于通过单DNA微操作来研究DNA-蛋白质复合体。利用磁钳和微吸管操纵附着在微米级颗粒上的DNA，将测量核蛋白复合体的形成动力学、机械性能和催化功能。这些蛋白质-DNA复合体的自组装、活性和拆解将通过精确的时间分辨的机械性能测量来研究，即分子长度作为作用力的函数。这种简单而通用的方法可以观察重组DNA结构的蛋白质的反应动力学和平衡状态。该项目还将包括开发横向磁性镊子方法，包括同时对两个DNA片段进行实验的技术。这项研究的更广泛影响将通过其高度跨学科的性质，以及通过开发用于研究DNA-蛋白质相互作用的新生物物理技术(横向磁性镊子和组合磁性镊子/微吸管方法)来实现。与此同时，拟议的活动将侧重于博士和博士后的研究教育，将物理学和生物学结合起来。该奖项的资金由数学和物理科学局的物理司和多学科活动办公室以及生物科学局的分子和细胞生物科学司提供。

This project focuses on the study of DNA-protein complexes through single-DNA micromanipulation. Using magnetic tweezers and micropipette manipulation of DNAs attached to micron-sized particles, the dynamics of formation, mechanical properties, and catalytic function of nucleoprotein complexes will be measured. Self-assembly, activities and disassembly of these protein-DNA complexes will be studied via precise time-resolved measurement of mechanical properties, i.e. molecular lengths as a function of applied force. This simple yet versatile approach allows observation of kinetics of reactions as well as equilibrium states, for proteins that reorganize DNA structure. The project will also include development of transverse magnetic tweezer methods, including techniques for experiments on two DNA segments at once. Broader impacts of this research will be via its highly interdisciplinary nature, and by development of new biophysical techniques (transverse magnetic tweezer and combined magnetic tweezer/micropipette methods) for study of DNA-protein interactions. Along with this, the proposed activities will focus on research education for Ph.D. and postdoctoral students, combining physics and biology. Funding for this award is provided by the Division of Physics and the Office of Multidisciplinary Activities in the Mathematical and Physical Sciences Directorate and by the Division of Molecular and Cellular Biosciences in the Biosciences Directorate.