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

使用单 DNA 显微操作研究核蛋白复合物的自组织、分子力学和催化功能

• 批准号: 0445565
• 负责人: John Marko
• 金额: $ 48.89万
• 依托单位: University of Illinois at Chicago
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-01 至 2008-10-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0445565&HistoricalAwards=false
• 关键词: Self; organization; molecular; mechanics; catalytic

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.

本项目主要通过单dna显微操作研究dna -蛋白复合物。利用磁镊子和微移液管操作附着在微米大小颗粒上的dna，将测量核蛋白复合物的形成动力学、力学性质和催化功能。这些蛋白质- dna复合物的自组装、活性和拆卸将通过精确的时间分辨机械性能测量来研究，即分子长度作为施加力的函数。这种简单而通用的方法允许观察反应动力学以及平衡状态，用于重组DNA结构的蛋白质。该项目还将包括横向磁镊子方法的开发，包括同时对两个DNA片段进行实验的技术。这项研究的广泛影响将通过其高度跨学科的性质，以及通过开发新的生物物理技术（横向磁镊子和组合磁镊子/微移液管方法）来研究dna -蛋白质相互作用。与此同时，计划的活动将集中在物理学和生物学相结合的博士和博士后的研究教育上。该奖项的资金由数学和物理科学理事会的物理司和多学科活动办公室以及生物科学理事会的分子和细胞生物科学司提供。