DNA damage response and repair of a broken chromosome

DNA 损伤反应和断裂染色体的修复

• 批准号: 10387373
• 负责人: JAMES E HABER
• 金额: $ 17.16万
• 依托单位: BRANDEIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-06-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10387373
• 关键词: Affect; Animal Model; Area; Autophagocytosis; Biochemistry; Biological Models; Cell Cycle Arrest; Cell Cycle Progression; Cells; Chromatin; Chromosomes; Complex; DNA Damage; DNA Repair; DNA damage checkpoint; Double Strand Break Repair; Event; Funding; Genes; Genetic Recombination; Genomic Instability; Goals; Hand; Human; Malignant Neoplasms; Mammalian Cell; Molecular; Monitor; Mutation; National Institute of General Medical Sciences; Process; Research; Saccharomycetales; Signal Transduction; Single-Stranded DNA; Site; base; cancer cell; homologous recombination; in vivo; insight; repaired; response

Project Summary The overall goal of NIGMS-funded research in my lab is to describe the molecular and cellular mechanisms by which cells sense the presence of DNA damage and carry our repair of chromosomal double-strand breaks (DSBs). Using budding yeast as a model system, it is possible to induce site-specific DSBs with a high degree of synchrony not yet possible in mammalian cells, allowing “in vivo biochemistry” approaches to monitor intermediate steps in DSB repair and DNA damage signaling. The first of three foci of this proposal investigates key questions concerning DSB repair by homologous recombination. How are homologous donor sequences found and used to repair a DSB? How are mismatches tolerated and repaired during different steps of recombination? How do cells deal with chromatin during repair and how is chromatin re- established after repair is complete? And how is gene editing accomplished using single-stranded DNA templates? The second area seeks to understand what is the basis of the 1000-fold increase in mutations associated with DSB repair and how microhomologies are used in repair-dependent template switching, creating complex chromosome rearrangements analogous to events recently found in human cancers. The third main objective is to understand how the DNA damage checkpoint and DNA damage-induced autophagy are regulated. We wish to determine: How does the DNA damage response affect DSB repair? How is the DNA damage checkpoint maintained and how is autophosphorylation of Mec1ATR regulated? What is the basis of a regulatory hand-off between the damage response and the spindle assembly checkpoint? Finally, we will determine the targets of DNA damage-induced autophagy and how autophagy contributes to cell cycle arrest in response to even a single unrepaired DSB. These studies will provide new insights and guidance in defining the DSB repair and checkpoint signaling in human cells.

项目摘要 在我的实验室里，NIGMS资助的研究的总体目标是描述 细胞感知DNA损伤的存在并进行修复的机制。 染色体双链断裂（DSB）。使用芽殖酵母作为模型系统， 为了诱导具有在哺乳动物细胞中尚不可能的高度同步性的位点特异性DSB， 允许“体内生物化学”方法监测DSB修复和DNA修复中的中间步骤， 损坏信号本建议的三个重点中的第一个重点调查了以下关键问题： 通过同源重组进行DSB修复。如何找到同源供体序列， 用来修理DSB的如何在不同的步骤中容忍和修复失配 重组？细胞在修复过程中如何处理染色质，染色质是如何修复的？ 修复完成后，？基因编辑是如何使用单链 DNA模板？第二个领域试图理解1000倍增长的基础是什么 与DSB修复相关的突变以及微同源性如何用于修复依赖性 模板转换，产生复杂的染色体重排，类似于最近发生的事件 在人类癌症中发现。第三个主要目标是了解DNA损伤 检查点和DNA损伤诱导的自噬受到调节。我们希望确定： DNA损伤反应影响DSB修复？DNA损伤检查点是如何维持的 Mec1ATR的自磷酸化是如何调节的？监管放手的依据是什么 损坏反应和主轴装配检查点之间的联系最后，我们将确定 DNA损伤诱导的自噬的靶点以及自噬如何促进细胞周期停滞 即使是一个未修复的DSB。这些研究将提供新的见解， 指导定义DSB修复和人类细胞中的检查点信号传导。