DNA Damage & DNA Replication: a Complex Relationship

DNA损伤

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要我们的总体目标是： i. 定义 Mre11 复合体激活 ATM 的机制；定义 Mre11 复合物在 DNA 复制中的作用； iii. 定义 Mre11 复合物在 DNA 复制中的作用； DNA 复制压力的响应作为 DNA 复制压力重点的一部分，我们的重点包括 RTEL1 的作用是减轻端粒的复制压力，这是酵母和小鼠遗传学的结合。我们之前在这些主题上的工作采用生物化学和结构生物学来解决这些问题。带来了与人类健康相关的重大科学进步和见解。询问的内容有： • 酵母遗传筛选和癌症基因组数据挖掘揭示了 Rad50 功能的分离影响 ATM 依赖性 DNA 损伤信号传导而不影响 DNA 修复功能的突变这些突变构成了破译其机制的资源。 Mre11 复合物激活 ATM 激酶以启动 DNA 损伤信号传导。 • 我们在小鼠中模拟了三种复发性肿瘤携带的突变，这些突变被证实是这些动物模型具有独特的 ATM 激活能力，但擅长 DNA 修复。描述 Mre11 复合体依赖的 ATM 功能的机会。 • 我们已经确定了在 DNA 复制叉上发挥作用的因素，其作用方式取决于 Mre11 复合体的一个重要重点是了解它们的功能作用。促进DNA精确复制的因素的缺陷与人类高度相关。疾病，因此了解这一基本过程是我们工作的一个重要优先事项。 •Mre11 复合体在复制叉上发挥作用，而 RTEL1 是一种解旋酶，可促进我们发现 RTEL1 影响端粒 DNA 的丰度和复制。称为 TERRA 的长非编码 RNA 的配置，从亚端粒区域转录我们在这方面工作的目标有两个：首先，了解 RTEL1 的作用。其次，利用这些信息来阐明端粒的功能。 TERRA，这是一个长期存在的问题。