Molecular Studies Of Protein-DNA Interactions

蛋白质-DNA 相互作用的分子研究

• 批准号: 7593637
• 负责人: Peggy Hsieh
• 金额: $ 41.08万
• 依托单位: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7593637
• 关键词: Aging; Atomic Force Microscopy; Binding; Biochemical; Biological; Cell physiology; Cells; Complex; Condition; DNA Damage; DNA Repair; DNA Repair Pathway; DNA lesion; DNA repair protein; DNA-Protein Interaction; Genome Stability; Genotoxic Stress; Goals; Hereditary Malignant Neoplasm; Immunofluorescence Microscopy; In Vitro; Light; Malignant Neoplasms; Mammalian Cell; Molecular; Monitor; Numbers; Organism; Pathway interactions; Post-Translational Protein Processing; Premature aging syndrome; Protein Array; Proteins; Research; Signal Pathway; Surface Plasmon Resonance; Time; mutant; protein protein interaction; repaired; response; single molecule; trafficking

Using a variety of biochemical, structural, and molecular and cell biological approaches, we are studying the molecular mechanisms underlying the diverse array of cellular functions involving DNA repair proteins. Surface plasmon resonance studies allow us to monitor in real-time key protein-protein and protein-DNA interactions formed in vitro under a variety of conditions to ask about complex stability and to characterize various aspects of complex formation utilizing both wild type and mutant proteins. Fluorescently tagged proteins and immunofluorescence microscopy are being used to study protein-protein interactions as well as traffic within different cell compartments. Modulation of the levels of repair proteins, post-translational modifications, and subcellular localization are being examined in response to a variety of genotoxic stresses. Single molecule studies using atomic force microscopy allow us to visualize multi-protein complexes bound to DNA lesions. These studies can help shed light on specific DNA repair pathways as well as general cellular responses to DNA damage that may be relevant to cancer and aging.

使用各种生物化学，结构，分子和细胞生物学方法，我们正在研究涉及DNA修复蛋白的细胞功能的不同阵列的分子机制。 表面等离子体共振研究使我们能够实时监测在各种条件下体外形成的关键蛋白质-蛋白质和蛋白质-DNA相互作用，以询问复合物的稳定性，并利用野生型和突变蛋白质表征复合物形成的各个方面。 标记蛋白质和免疫荧光显微镜正被用于研究蛋白质-蛋白质相互作用以及不同细胞隔室内的交通。 调节修复蛋白的水平，翻译后修饰，和亚细胞定位正在检查在响应各种遗传毒性应力。 使用原子力显微镜的单分子研究使我们能够可视化与DNA损伤结合的多蛋白质复合物。 这些研究可以帮助阐明特定的DNA修复途径以及可能与癌症和衰老相关的DNA损伤的一般细胞反应。

项目成果

Determination of protein-DNA binding constants and specificities from statistical analyses of single molecules: MutS-DNA interactions.

• DOI: 10.1093/nar/gki708
• 发表时间: 2005
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Yang Y;Sass LE;Du C;Hsieh P;Erie DA
• 通讯作者: Erie DA

Requirement for Phe36 for DNA binding and mismatch repair by Escherichia coli MutS protein.

大肠杆菌 MutS 蛋白 DNA 结合和错配修复需要 Phe36。

• DOI: 10.1093/nar/28.18.3564
• 发表时间: 2000
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Yamamoto,A;Schofield,MJ;Biswas,I;Hsieh,P
• 通讯作者: Hsieh,P