An AFM Study of DNA Damage and Repair
DNA 损伤与修复的 AFM 研究
基本信息
- 批准号:0450835
- 负责人:
- 金额:$ 48万
- 依托单位:
- 依托单位国家:美国
- 项目类别:Continuing Grant
- 财政年份:2005
- 资助国家:美国
- 起止时间:2005-02-01 至 2009-01-31
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
The purpose of this research is to further the understanding of DNA damage and repair by elucidating the relationship between these processes and DNA nanomechanics and by visualizing repair protein activities on DNA using atomic force microscopy techniques. The objectives are: 1) optimize the AFM platform for single-molecule DNA measurements 2) examine various lesions in individual DNA molecules and follow in the AFM their direct reversal by DNA repair enzymes 3) use AFM imaging and force spectroscopy to visualize the mismatch repair reaction of E. coli. To achieve these objectives the AFM platform needs to be optimized to reduce force errors to single piconewtons and to accelerate image acquisition by using the ultra-small cantilever technology. DNA repair proteins such as photolyases, endonucleases and ligases will be used as damage markers and will be imaged in complexes with DNA by AFM in order to locate and identify the damage sites. Force spectroscopy measurements will determine the mechanical fingerprints of various types of DNA damage caused by UV and gamma radiation and will directly follow damage reversal in the presence of the repair activities. This project will also examine and visualize the DNA mismatch repair reaction using E. coli repair activities and AFM imaging and force spectroscopy technologies. This project will provide an exciting education and research opportunity for three graduate students at the interface between engineering and biology and will likely develop new ultra sensitive assays for DNA damage detection that is of significance to many areas of biology.
本研究的目的是通过阐明这些过程与DNA纳米力学之间的关系,并通过使用原子力显微镜技术可视化DNA上的修复蛋白活性,进一步了解DNA损伤和修复。目标是:1)优化单分子DNA测量的AFM平台;2)检查单个DNA分子中的各种病变,并在AFM中通过DNA修复酶直接逆转它们;3)使用AFM成像和力谱可视化大肠杆菌错配修复反应。为了实现这些目标,AFM平台需要进行优化,以将力误差降低到单皮牛顿,并通过使用超小型悬臂技术加速图像采集。DNA修复蛋白如光解酶、内切酶和连接酶将被用作损伤标记物,并将在与DNA的复合物中通过AFM成像以定位和识别损伤位点。力谱测量将确定由紫外线和伽马辐射引起的各种类型DNA损伤的机械指纹,并将直接跟踪修复活动中存在的损伤逆转。该项目还将使用大肠杆菌修复活性、AFM成像和力谱技术来检测和可视化DNA错配修复反应。该项目将为工程学和生物学交叉领域的三名研究生提供令人兴奋的教育和研究机会,并可能开发出对生物学许多领域具有重要意义的新型超灵敏DNA损伤检测方法。
项目成果
期刊论文数量(0)
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会议论文数量(0)
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Piotr Marszalek其他文献
Accurate Refolding of Experimentally Determined Protein Mechanical Unfolding Intermediates via All-Atom Molecular Dynamics Simulations
- DOI:
10.1016/j.bpj.2017.11.2874 - 发表时间:
2018-02-02 - 期刊:
- 影响因子:
- 作者:
David Wang;Piotr Marszalek - 通讯作者:
Piotr Marszalek
Full Reconstruction of a Vectorial Protein Folding Pathway by Afm and Smd: Insights Into the Co-Translational Folding of the Nascent-Polypeptide-Chain
- DOI:
10.1016/j.bpj.2010.12.2829 - 发表时间:
2011-02-02 - 期刊:
- 影响因子:
- 作者:
Whasil Lee;Xiancheng Zeng;Huan-Xiang Zhou;Vann Bennett;Weitao Yang;Piotr Marszalek - 通讯作者:
Piotr Marszalek
Piotr Marszalek的其他文献
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{{ truncateString('Piotr Marszalek', 18)}}的其他基金
EAGER: Exploring the Quantum-Mechanical Basis of Odorant Detection by Olfactory Receptors
EAGER:探索嗅觉受体气味检测的量子力学基础
- 批准号:
2105612 - 财政年份:2021
- 资助金额:
$ 48万 - 项目类别:
Standard Grant
Transition To Excellence: From Single-Molecule Force Spectroscopy to Single-Particle Cryogenic Electron Microscopy
向卓越过渡:从单分子力谱到单粒子低温电子显微镜
- 批准号:
2118357 - 财政年份:2021
- 资助金额:
$ 48万 - 项目类别:
Standard Grant
Workshop: Progress and Prospects of Single Molecule Force Spectroscopy in Biological and Chemical Sciences Workshop; May 30 - June 2, 2019; Durham, North Carolina
研讨会:单分子力谱在生物化学科学中的进展与展望研讨会;
- 批准号:
1856726 - 财政年份:2019
- 资助金额:
$ 48万 - 项目类别:
Standard Grant
Molecular Mechanisms of Spontaneous and Hsp 70-assisted Renaturation of Misfolded Proteins
错误折叠蛋白自发复性和 Hsp 70 辅助复性的分子机制
- 批准号:
1817556 - 财政年份:2018
- 资助金额:
$ 48万 - 项目类别:
Standard Grant
Vectorial Folding of Large, Multidomain Proteins
大型多域蛋白质的矢量折叠
- 批准号:
1517245 - 财政年份:2015
- 资助金额:
$ 48万 - 项目类别:
Continuing Grant
Investigating DNA Mismatch Repair Through Single-Molecule Approaches
通过单分子方法研究 DNA 错配修复
- 批准号:
1244297 - 财政年份:2013
- 资助金额:
$ 48万 - 项目类别:
Continuing Grant
Vectorial Folding of Proteins and Nascent Polypeptide Chains by AFM and Computer Simulations
通过 AFM 和计算机模拟进行蛋白质和新生多肽链的矢量折叠
- 批准号:
1052208 - 财政年份:2011
- 资助金额:
$ 48万 - 项目类别:
Continuing Grant
Investigating Conformations of Single Polysaccharides and Nucleic Acids by Force Spectroscopy
通过力谱研究单多糖和核酸的构象
- 批准号:
0717770 - 财政年份:2007
- 资助金额:
$ 48万 - 项目类别:
Continuing Grant
Force-induced Conformational Transitions in Single Polysaccharide Molecules by AFM
通过 AFM 力诱导单多糖分子的构象转变
- 批准号:
0243360 - 财政年份:2002
- 资助金额:
$ 48万 - 项目类别:
Continuing Grant
Force-induced Conformational Transitions in Single Polysaccharide Molecules by AFM
通过 AFM 力诱导单多糖分子的构象转变
- 批准号:
0110093 - 财政年份:2001
- 资助金额:
$ 48万 - 项目类别:
Continuing Grant
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