DNA Damage & DNA Replication: a Complex Relationship

DNA损伤

• 批准号: 10449117
• 负责人: John HJ Petrini
• 金额: $ 94.96万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10449117
• 关键词: ATM activation; ATM function; Address; Affect; Animal Model; Area; Biochemistry; Bone marrow failure; Chromosomal Instability; Complex; DNA; DNA Damage; DNA Repair; DNA Replication Damage; DNA biosynthesis; DNA replication fork; Defect; Development; Eukaryota; Genetic Screening; Genome Stability; Genomic Instability; Goals; Health; Human; Human Chromosomes; Immune; Laboratories; Light; Maintenance; Malignant Neoplasms; Mining; Modeling; Mus; Mutation; Neurologic; Process; Proteins; Recurrent tumor; Resources; Risk; Role; Signal Transduction; Syndrome; Untranslated RNA; Work; Yeasts; ataxia telangiectasia mutated protein; cancer genomics; experimental study; genomic data; helicase; human disease; insight; mouse genetics; repair function; replication stress; reproductive; response; structural biology; telomere; yeast genetics

PROJECT SUMMARY/ABSTRACT Our overarching goals are: i. to define the mechanism(s) of ATM activation by the Mre11 complex; ii. to define the role(s) of the Mre11 complex in DNA replication; iii. To define the role(s) of the Mre11 complex in response to DNA replication stress. As part of the emphasis of DNA replication stress, our focus includes RTEL1 which acts to mitigate replication stress at the telomere. A combination of yeast and mouse genetics, biochemistry, and structural biology are employed to address these issues. Our previous work on these topics led to substantial scientific progress and insight regarding processes relevant to human health. Specific areas of inquiry are: • Genetic screens in yeast and mining of cancer genomic data revealed separation of function Rad50 mutations that affect ATM dependent DNA damage signaling without affecting the DNA repair functions of the Mre11 complex. These mutations constitute a resource for deciphering the mechanism by which the Mre11 complex activates the ATM kinase to initiate DNA damage signaling. • We have modeled three recurrent tumor borne mutations in mice that were confirmed to be hypomorphic for ATM activation but proficient in DNA repair. These animal models offer a unique opportunity to delineate Mre11 complex-dependent ATM functions. • We have identified factors that function at the DNA replication fork in a manner that depends on the Mre11 complex. An important focus of our laboratory is to understand the functional role(s) of those factors. Defects in factors that promote accurate DNA replication are highly correlated with human disease, and so understanding this fundamental process is an important priority in our work. • Whereas the Mre11 complex functions at the replication fork, RTEL1 is a helicase that promotes accurate replication of telomeric DNA. We have discovered that RTEL1 influences the abundance and disposition of a long non coding RNA, called TERRA, that is transcribed from the subtelomeric regions of all eukaryotes. Our goal in this aspect of our work is two fold. First, to understand the role of RTEL1 in maintaining telomere stability. Second, to use that information to shed light on the function of TERRA, which is a long standing question.

项目总结/摘要 我们的首要目标是：i.定义Mre 11复合物激活ATM的机制; ii.到 定义Mre 11复合物在DNA复制中的作用; iii.要定义Mre 11复合体在 DNA复制应激反应。作为DNA复制压力的重点的一部分，我们的重点包括 RTEL 1，其作用是减轻端粒处的复制应激。酵母和老鼠基因的结合， 生物化学和结构生物学被用来解决这些问题。我们以前在这些主题上的工作 在与人类健康有关的过程方面取得了重大的科学进展和深刻的见解。具体领域 调查是： ·酵母中的遗传筛选和癌症基因组数据的挖掘揭示了Rad 50功能的分离 影响ATM依赖性DNA损伤信号而不影响DNA修复功能的突变 Mre 11复合体这些突变构成了一个资源，用于破译的机制， Mre 11复合物激活ATM激酶以启动DNA损伤信号传导。 ·我们在小鼠中建立了三种复发性肿瘤突变模型，这些突变已被证实是 ATM激活亚型但精通DNA修复。这些动物模型提供了一个独特的 描述Mre 11复杂依赖的ATM功能的机会。 ·我们已经确定了在DNA复制叉上起作用的因素，其作用方式取决于DNA复制叉的大小。 Mre 11复合体。我们实验室的一个重要重点是了解那些 因素促进准确DNA复制的因子的缺陷与人类的遗传高度相关。 因此，了解这一基本过程是我们工作中的一个重要优先事项。 ·尽管Mre 11复合物在复制叉处起作用，但RTEL 1是一种解旋酶， 端粒DNA的精确复制。我们已经发现RTEL 1影响了 一个长的非编码RNA，称为TERRA，是从亚端粒区域转录的 所有的真核生物。我们在这方面的工作有两个目标。首先，了解RTEL 1的作用 维持端粒的稳定性。第二，利用这些信息来阐明 这是一个长期存在的问题。