Single-Molecule Assembly of Protein-DNA Complexes

蛋白质-DNA 复合物的单分子组装

基本信息

批准号：
9276464
负责人：
Stephen Charles Kowalczykowski
金额：
$ 9.97万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-07-15 至 2020-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9276464
关键词：
Address Affect Area Behavior Biochemical Biological Biological Process Buffers Bypass Cell physiology Chromosomal translocation Complement Complex DNA DNA Damage DNA Polymerase beta DNA Repair DNA biosynthesis DNA-Directed RNA Polymerase DNA-Protein Interaction Detection Ensure Enzymes Event Filament Fluorescence Microscopy Genome Stability Genomic Instability Health Heterogeneity Human Image Imagery Individual Kinetics Laboratories Lead Lesion Light Maintenance Measures Mediator of activation protein Methods Molecular Monitor Motor Mutation Nucleoproteins Nucleosomes Positioning Attribute Process Property Proteins Regulation Research Site Slide Speed Structure Time chromatin remodeling ds-DNA genetic information insight instrument instrumentation interest new technology protein phosphatase inhibitor-2 single molecule success transmission process

项目摘要

DESCRIPTION (provided by applicant): The broad objective of this proposal is to visualize and quantify the dynamic behavior of biological processes at the single-molecule level. We have developed new experimental strategies for studying protein-DNA interactions at the single-molecule level. We devised several approaches wherein we could easily manipulate an individual DNA molecule and expose it to a variety of different proteins, enzymes, and buffers. Using fluorescent detection, we could image individual proteins on DNA or detect their action on individual DNA molecules, typically in real-time. We propose to continue to use this approach to study unresolved aspects of recombinational DNA repair, chromatin remodeling, and DNA replication. We seek to continue to address unresolved questions regarding the assembly and disassembly of RecA/Rad51 nucleoprotein filaments, their control and function in homology-directed DNA repair. In addition, we propose to expand our interests and success in the area of chromatin remodeling by motor proteins. And finally, we have initiated studies on single-molecule imaging of DNA replication, and propose to address important issues that intersect with recombinational DNA repair.

描述（由适用提供）：该建议的广泛目标是可视化和量化单分子水平生物过程的动态行为。我们开发了新的实验策略，用于研究单分子水平的蛋白-DNA相互作用。我们设计了几种方法，其中我们可以轻松地操纵单个DNA分子，并将其暴露于各种不同的蛋白质，酶，并使用荧光检测，我们可以在DNA上对单个蛋白进行成像或检测其对单个DNA分子的作用，通常是实时实时的。我们建议继续使用这种方法来研究重组DNA修复，染色质重塑和DNA复制的未解决方面。我们试图继续解决有关RECA/RAD51核蛋白丝的组装和拆卸的尚未解决的问题，它们在同源指导的DNA修复中的控制和功能。此外，我们建议扩大运动蛋白重塑染色质重塑领域的兴趣和成功。最后，我们启动了有关DNA复制的单分子成像的研究，并提出了解决与重组DNA修复相交的重要问题的建议。