The role of chromatin remodeling factors in DNA double strand break repair

染色质重塑因子在DNA双链断裂修复中的作用

基本信息

批准号：
10201222
负责人：
Renee Bouley
金额：
$ 17万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10201222
关键词：
Affect Biochemical Biological Assay Catalogs Cells Chemicals Chromatin Chromatin Remodeling Factor Chromosomal Instability Chromosomal Stability Chromosome Deletion Chromosome abnormality Chromosome inversion Chromosomes Complex Copy Number Polymorphism DNA DNA Damage DNA Double Strand Break DNA Repair DNA damage checkpoint DNA lesion DNA replication fork Data Databases Deposition Double Strand Break Repair Eukaryota Excision Failure Fission Yeast Genes Genetic Recombination Genomic Instability Goals HTATIP gene Histones Homologous Gene Human Malignant Neoplasms Modeling Molecular Chaperones Mutate Mutation Nucleosomes Observational Study Pathway interactions Play Process Property Proteins RAD52 gene Rad51 recombinase Repair Complex Role Sister Somatic Mutation System Testing Work Yeasts cancer cell cancer genome chromatin remodeling gene repression genetic analysis histone acetyltransferase homologous recombination human disease metaplastic cell transformation molecular modeling mutant novel prevent protein function protein protein interaction repaired replication stress tumor tumorigenesis yeast two hybrid system

项目摘要

7. PROJECT SUMMARY/ABSTRACT Accurate DNA double strand break (DSB) repair is important to maintain chromosomal stability. In the absence of DSB repair, translocations, deletions, duplications and inversions of chromosomal DNA may occur that can drive tumorigenesis. Homologous recombination (HR) is the major mechanism for DSB repair and is highly conserved across eukaryotes from yeast to human. HR is generally error-free because it copies any missing information from an undamaged homologue or sister chromosome template. However, not all HR is error-free. A number of conditions, that includes replication stress from spontaneous, chemical or physical DNA damage, can activate error-prone recombination pathways that leads to intra-chromosomal deletions (ICDs) as well as other chromosome rearrangements. The central HR gene RAD52 participates in both error-free and error-prone repair. How RAD52 choses between these opposing pathways is poorly understood. It is our central hypothesis that chromatin remodeling factors are responsible for guiding this choice. Previous work utilizing a two-hybrid screen in the fission yeast Schizosaccharomyces pombe identified a physical interaction between RAD52 with the histone acetyltransferase KAT5 and the histone chaperone HIRA. KAT5 (also known as TIP60 in humans; Mst1 in yeast) is required to target chromatin for remodeling as well as DNA damage checkpoint activation. HIRA (Hip1 in yeast) functions in establishing centromeric chromatin, targeted transcriptional repression and replication-independent histone assembly. Our preliminary data demonstrate that KAT5 channels DSBs toward error-free repair while HIRA directs DSBs toward error-prone repair that results in ICDs. The goal of this proposal is to understand how the physical interactions and chromatin remodeling properties of KAT5 and HIRA impact the decision by RAD52 to function in error-free versus error-prone repair. We propose two Specific Aims: In Aim 1 we will perform comprehensive protein modeling to predict specific residues involved in the interactions between KAT5, HIRA and RAD52. These residues will be altered and examined for repair pathway choice using well-defined chromosomal recombination assays. In Aim 2 we will examine the effect of KAT5, HIRA and RAD52 mutations found in human tumors for their ability to influence error-free and/or error-prone repair pathway choice. These studies will determine the role of chromatin remodeling in accurate DNA damage repair that is required to prevent genetic alterations that leads to cancer.

7。项目摘要/摘要准确的DNA双链断裂（DSB）修复对于维持染色体稳定性很重要。在缺席的情况下可能会发生DSB修复，易位，缺失，重复和反转染色体DNA 驱动肿瘤发生。同源重组（HR）是DSB修复的主要机制，高度从酵母到人类的真核生物遍布整个真核生物。人力资源通常是没有错误的，因为它复制了任何丢失的来自未损坏的同源物或姐妹染色体模板的信息。但是，并非所有人力资源都没有错误。许多条件，包括自发，化学或物理DNA损伤的复制应力，可以激活容易出错的重组途径，从而导致染色体内缺失（ICD）以及其他染色体重排。中央人力资源基因RAD52参与无错误和容易出错的参与维修。 RAD52如何在这些相反的途径之间选择不理解。这是我们的中心假设染色质重塑因素负责指导此选择。以前利用裂变酵母酸果实糖果疗中的两个杂交屏幕Pombe确定了一个物理的工作 Rad52与组蛋白乙酰转移酶Kat5和组蛋白伴侣HIRA之间的相互作用。 Kat5 （在人类中也称为tip60；酵母中的Mst1）才能靶向染色质进行重塑和DNA 损坏检查点激活。 HIRA（酵母中的HIP1）在建立焦糖染色质中起作用，靶向转录抑制和无关的组蛋白组装。我们的初步数据表明 KAT5通道DSB朝着无错误维修时 ICD。该建议的目的是了解物理互动和染色质重塑 KAT5和HIRA的属性会影响Rad52在无错误和容易出错的修复中起作用的决定。我们提出了两个具体目标：在AIM 1中，我们将执行全面的蛋白质建模，以预测相互作用中涉及的特定残基在Kat5，Hira和Rad52之间。这些残留物将更改并检查使用修复途径的选择定义明确的染色体重组测定法。在AIM 2中，我们将检查Kat5，Hira和Rad52的效果在人类肿瘤中发现的突变具有影响无错误和/或容易发生修复途径的能力。这些研究将确定染色质重塑在需要的准确DNA损伤修复中的作用为了防止导致癌症的遗传改变。