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：
8900314
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 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8900314
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必不可少的调节周期事件。每个细胞周期，必须选择并激活数千个复制起始位点（起源），以确保基因组精确地复制在S期范围内。无法正确调节DNA复制程序可能会导致灾难性的基因组不稳定性。我们在D. Melanogaster和S. cerevisiae中的工作确定了染色质结构和ATP依赖性染色质重塑活性是原点选择的重要决定因素。我们正在提出遗传，基因组和生化方法，以了解局部染色质结构和ATP依赖性染色质重塑如何有助于选择和调节酵母和果蝇中复制起源。具体而言，我们将评估染色质动力学（独立的组蛋白H3/H4交换）与原点选择和激活之间的相互作用。我们还将使用全面的基因组方法来测试候选ATP染色质重塑活动在起源选择和激活中的作用。我们最近开发了一个全基因组的“足迹”测定法，该测定法将用于解决有关起源体系结构和关键复制起始因子的分布的基本问题。作为我们的Modencode研究的一部分，我们确定了NURF染色质重塑复合物的多个成分是对ORC定位具有强的预测值。我们将测试NURF在指定果蝇复制起源中的作用。通过利用酵母和果蝇的实验强度，我们将对染色质结构如何指定复制的真核起源产生重要的机理见解。