Deciphering the chromatin-based DNA damage response pathway

破译基于染色质的 DNA 损伤反应途径

• 批准号: 10759124
• 负责人: Justin Wai Chung Leung
• 金额: $ 37.54万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10759124
• 关键词: Deciphering; chromatin; based; DNA; damage

PROJECT SUMMARY DNA damage is a constant threat to our genome. Our body has evolved a surveillance mechanism, namely the DNA damage response (DDR) pathway, to protect our cells from genotoxic insults. Defective DDR pathway leads to unfaithful repair of DNA breaks which results in accumulation of mutations, chromosome rearrangement, and genome instability. Although much is known about how chromatin senses and responds to DNA damage, a significant gap in knowledge exists with the poorly characterized mechanism by which damaged chromatin transduces the signal to recruit effector proteins to DNA breaks. Such knowledge is imperative to understand how cells precisely execute the correct pathway by recruiting the right repair protein complex in a temporal and spatial manner which is essential for maintaining our genome integrity. This research program aims to address this fundamental question in the chromatin-based DDR pathway. Our expertise lies in genome editing, functional proteomics, molecular biology, cell biology, and biochemistry. We combine our techniques of protein purification, confocal microscopy, laser-induced single cell micro-irradiation, cell-based reporter assays, protein tagging, and quantitative mass spectrometry for an overall multidisciplinary approach. The goals of this MIRA project are to obtain a comprehensive molecular understanding of the chromatin-based DDR pathway by 1) determining the epigenetic profile in the DDR pathway, 2) characterizing the functions of the novel DNA repair proteins and 3) elucidating the mechanism of how damaged chromatin orchestrates different DNA repair pathways during the cell cycle in the context of damaged chromatin. Using fractionation purification-coupled proteomic approach and cell-based functional assays, we have identified more than twenty novel DNA repair proteins. Our long-term goals are to build a physical and genetic network within the chromatin-based DDR pathway and understand its impact in human health. Overall, these studies will open up a new arena for the DNA repair field, provide insight into the etiology of cancer and genome instability-related genetic diseases that will lay the foundation for translational research and therapeutic strategy development.

项目摘要 DNA损伤是对我们基因组的持续威胁。我们的身体已经进化出一种监视机制，即 DNA损伤反应（DDR）途径，以保护我们的细胞免受遗传毒性的伤害。DDR通路电极导线存在缺陷 DNA断裂的不忠实修复导致突变的积累，染色体重排， 基因组不稳定性虽然我们对染色质如何感知和响应DNA损伤已经有了很多了解， 由于染色质受损的机制特征不清，目前的知识存在显着差距 转导信号以募集效应蛋白到DNA断裂处。这些知识是理解 细胞如何精确地执行正确的途径，通过招募正确的修复蛋白复合物， 空间方式对于维持我们基因组的完整性至关重要。该研究计划旨在解决 这是基于染色质的DDR途径中的基本问题。我们的专长在于基因组编辑，功能性 蛋白质组学、分子生物学、细胞生物学和生物化学。我们联合收割机将蛋白质纯化技术， 共聚焦显微镜，激光诱导的单细胞微辐射，基于细胞的报告基因测定，蛋白质标记，和 定量质谱法用于整体多学科方法。这个MIRA项目的目标是 通过以下方式获得对基于染色质的DDR途径的全面分子理解：1）确定 DDR途径中的表观遗传谱，2）表征新型DNA修复蛋白的功能和3） 阐明了受损的染色质如何协调不同的DNA修复途径的机制， 在染色质受损的情况下的细胞周期。采用分离纯化偶联蛋白质组学方法， 通过基于细胞的功能测定，我们已经鉴定了20多种新的DNA修复蛋白。我们的长期 目标是在基于染色质的DDR途径中建立物理和遗传网络并了解其 对人类健康的影响。总体而言，这些研究将为DNA修复领域开辟一个新的竞技场， 癌症和基因组不稳定性相关的遗传疾病的病因学，这将为 转化研究和治疗策略开发。