Chromatin architecture defines DNA replication origins

染色质结构定义了 DNA 复制起点

基本信息

批准号：
9113031
负责人：
David M MacAlpine
金额：
$ 29.39万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-08-09 至 2018-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9113031
关键词：
Address Architecture Binding Binding Sites Biochemical Biochemical Genetics Biological Assay Biological Models Cancer Biology Cell Cycle Cell division Chromatin Chromatin Remodeling Factor Chromosome Fragile Sites Chromosomes Complex Conserved Sequence Copy Number Polymorphism DNA DNA biosynthesis DNA replication origin Development Drosophila genus Elements Ensure Environment Failure Gene Amplification Genetic Genetic Transcription Genome Genome Stability Genomic Instability Genomic approach Genomics Health Histone H3 ISWI Individual Lead Link Maps Mediating NURF Nuclear Nucleosomes Nucleotides Peptide Initiation Factors Positioning Attribute Pre-Replication Complex Predictive Value Process Protein Footprinting Regulation Replication Initiation Replication Origin Research Resolution Role S Phase Saccharomyces cerevisiae Site Specific qualifier value Staging Testing Time Tissues Trans-Activators Work Yeasts cell type chromatin remodeling cis acting element epigenome event cycle genetic approach genetic information genome-wide helicase insight origin recognition complex programs research study stem cell biology transcription factor tumorigenesis yeast genome

项目摘要

DESCRIPTION (provided by applicant): DNA replication is an essential and regulated cycle event required for the inheritance of genetic information. Every cell cycle, thousands of replication start sites (origins) must be selected and activated to ensure that the genome is copied exactly once within the confines of S-phase. Failure to properly regulate the DNA replication program may lead to catastrophic genomic instability. Our work in D. melanogaster and S. cerevisiae has identified chromatin architecture and ATP-dependent chromatin remodeling activities as important determinants of origin selection. We are proposing genetic, genomic and biochemical approaches to understand how the local chromatin architecture and ATP-dependent chromatin remodeling contribute to the selection and regulation of replication origins in yeast and Drosophila. Specifically, we will assess the interplay between chromatin dynamics (replication independent histone H3/H4 exchange) and origin selection and activation. We will also test the role of candidate ATP- dependent chromatin remodeling activities on origin selection and activation using comprehensive genomic approaches. We have recently developed a genome-wide 'footprinting' assay that will be used to address fundamental questions regarding origin architecture and the distribution of key replication initiation factors t individual origins. As part of our modENCODE studies, we identified multiple components of the NURF chromatin remodeling complex as having strong predictive value for ORC localization. We will test the role of NURF in specifying Drosophila origins of replication. By exploiting the experimental strengths of both yeast and Drosophila, we will generate important mechanistic insights into how the chromatin architecture specifies eukaryotic origins of replication.

描述（由申请人提供）：DNA复制是遗传信息遗传所需的重要且受调节的循环事件。每个细胞周期，必须选择并激活数千个复制起始位点（起点），以确保基因组在 S 期范围内准确复制一次。未能正确调节 DNA 复制程序可能会导致灾难性的基因组不稳定。我们对黑腹果蝇和酿酒酵母的研究已确定染色质结构和 ATP 依赖性染色质重塑活动是起源选择的重要决定因素。我们提出遗传、基因组和生化方法来了解局部染色质结构和 ATP 依赖性染色质重塑如何有助于酵母和果蝇复制起点的选择和调节。具体来说，我们将评估染色质动力学（复制独立的组蛋白 H3/H4 交换）与起源选择和激活之间的相互作用。我们还将使用综合基因组方法测试候选 ATP 依赖性染色质重塑活动对起源选择和激活的作用。我们最近开发了一种全基因组“足迹”测定，将用于解决有关起源结构和关键复制起始因子在各个起源上的分布的基本问题。作为 modENCODE 研究的一部分，我们发现 NURF 染色质重塑复合物的多个成分对 ORC 定位具有很强的预测价值。我们将测试 NURF 在指定果蝇复制起点中的作用。通过利用酵母和果蝇的实验优势，我们将对染色质结构如何指定真核复制起源产生重要的机制见解。