Regulation of DNA replication by histone modifications

通过组蛋白修饰调节 DNA 复制

• 批准号: 7268774
• 负责人: TOSHIO TSUKIYAMA
• 金额: $ 26.88万
• 依托单位: FRED HUTCHINSON CANCER RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7268774
• 关键词: Affect; Area; Biology; Cell Cycle; Cell Cycle Progression; Cell Cycle Stage; Cell Death; Cells; Chromatin; Chromatin Structure; Condition; DNA Repair; DNA biosynthesis; DNA replication origin; DNA-Directed DNA Polymerase; DNA-Protein Interaction; Genetic Transcription; Genome; Genomic Instability; Goals; Growth; Histones; Laboratories; Lead; Malignant Neoplasms; Mass Spectrum Analysis; Modification; Molecular; Monitor; Mutate; Mutation; Nuclear; Pattern; Play; Process; Regulation; Replication Origin; Role; Saccharomyces; Saccharomycetales; Stress; System; Testing; Time; base; cell growth; developmental disease; human disease; in vivo; instrument; novel

DESCRIPTION (provided by applicant): The broad, long term goal of the proposed study is to determine, at the molecular level, the mechanisms by which DNA replication occurs in vivo. Faithful duplication of the genome is an essential step in the cell cycle. Any disturbance in this process can lead to cell death, genomic instability, or unregulated cell growth. Therefore, studying the mechanisms of DNA replication will contribute not only to broad areas of basic biology, but also to understanding the molecular basis for human diseases, such as cancer and developmental disorders. DNA replication requires highly coordinated recruitment of replication factors to replication origins in a chromatin context, as well as progression of DNA polymerases through a chromatin template. It is therefore expected that histone modification patterns around replication origins strongly affect DNA replication. It is highly likely that proper histone modification patterns are established around replication origins before each DNA replication cycle. However, how histone modification patterns change around DNA replication origins during cell cycle progression and how specific patterns of histone modification affect DNA replication have not been determined, due to technical limitations. We have recently developed a new system to purify an autonomously replicating chromatin circle in microgram quantities. Combined with an ultra-sensitive mass spectrometry instrument, our system gives us a unique opportunity to identify histone modifications in an unbiased fashion around DNA replication origins during cell cycle progression. We will use our system to test our hypothesis that histone modifications are under dynamic regulation during cell cycle progression and play critical roles in DNA replication.

描述（由申请人提供）：拟议研究的广泛，长期目标是在分子水平上确定DNA在体内发生DNA复制的机制。基因组的忠实重复是细胞周期中的重要步骤。在此过程中的任何干扰都会导致细胞死亡，基因组不稳定性或不调节的细胞生长。因此，研究DNA复制的机制不仅会导致基本生物学的广泛领域，还将有助于了解人类疾病（例如癌症和发育障碍）的分子基础。 DNA复制需要高度协调的复制因子来复制因子在染色质环境中的复制起源，以及通过染色质模板的DNA聚合酶进展。因此，预计复制起源周围的组蛋白修饰模式强烈影响DNA复制。在每个DNA复制周期之前，围绕复制起源建立了适当的组蛋白修饰模式。然而，由于技术局限性，组蛋白的修饰模式在细胞周期进程过程中如何改变DNA复制起源周围的DNA复制起源以及组蛋白修饰的特定模式如何影响DNA复制。我们最近开发了一个新系统，以纯化自主复制染色质圆圈的微克量。与超敏感的质谱仪器相结合，我们的系统为我们提供了一个独特的机会，可以在细胞周期进程中以无偏见的形式鉴定组蛋白修饰。我们将使用系统来测试我们的假设，即在细胞周期进程中，组蛋白修饰受动态调节，并在DNA复制中起关键作用。