Comprehensive Characterization of Heterochromatin Domains

异染色质结构域的综合表征

• 批准号: 8424267
• 负责人: Aaron M. Johnson
• 金额: $ 23.64万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8424267
• 关键词: Address; Affect; Biology; Cell Extracts; Cells; Chromatin; Chromatin Modeling; Complex; Elements; Environment; Eukaryota; Event; Fission Yeast; Foundations; Functional RNA; Gene Expression; Gene Silencing; Genes; Genetic Transcription; Genome Stability; Genomics; Goals; Heterochromatin; Histone Deacetylase; Human; In Vitro; Lead; Maintenance; Mass Spectrum Analysis; Mediating; Modeling; Organism; Pathway interactions; Proteins; RNA; RNA Polymerase II; Recruitment Activity; Repression; Research Personnel; Saccharomycetales; Structure; System; Untranslated RNA; Yeast Model System; Yeasts; gene repression; histone modification; in vivo; prevent; protein complex; reconstitution

PROJECT SUMMARY/ABSTRACT Heterochromatin is a genomic structure that functions in all eukaryotes to determine cell identity and maintain genome stability by silencing genes. The requirements for heterochromatin assembly are incorporation into chromatin of silencing protein complexes with histone modification activity that lead to repressed gene expression. This general mechanism is conserved in all eukaryotes, though multiple varieties of heterochromatic domains exist, even within the same organism. Much is known about the factors that contribute to heterochromatin, yet many key events in specific pathways are poorly understood. One challenge in studying heterochromatin is the fact that manipulation of the central components in vivo often affects multiple types of heterochromatin or other related pathways in the cell. The aims of this project will address this problem in three ways: firstly, by using a functional in vitro reconstituted heterochromatin system to better understand the mechanism of heterochromatic silencing in an isolated environment. Secondly, using a pre- assembled heterochromatin domain to identify new factors within the cell that contribute to heterochromatin formation and maintenance. Third, by establishing a system to address the contribution of specific non-coding RNA elements to initiation and maintenance of heterochromatin. Initial studies in yeast models of heterochromatin will make important contributions to the field of chromatin biology and will also serve as a model for subsequent studies in the human system to be carried out after the transition to independent investigator. The primary goal of the project is to comprehensively characterize heterochromatin domains in order to compare conserved features that are central to gene silencing and genome stability. The diversity of human heterochromatic domains is evident, yet the general principles that govern assembly of these domains remain quite similar to those in the budding and fission yeast models. These basic similarities make it feasible to compare results of yeast studies to humans and also to use the approaches that have been developed in the simpler yeast models as a guide to study human heterochromatin. This theme is the foundation of this proposal and the direction I will take as I transition to independence.

项目摘要/摘要 异染色质是一种基因组结构，在所有真核生物中起作用，以确定细胞身份并维持 通过沉默基因进行基因组稳定性。异染色质组装的要求已纳入 带有组蛋白修饰活性的沉默蛋白复合物的染色质，导致抑制基因 表达。这种一般机制在所有真核生物中都保存，尽管多种多样 即使在同一生物体中，也存在异质域。对这些因素有很多了解 有助于异染色质，但是对特定途径中的许多关键事件却鲜为人知。一个挑战 在研究异染色质时，在体内操纵中心成分通常会影响多个 细胞中异染色质或其他相关途径的类型。该项目的目的将解决这个问题 以三种方式出现问题：首先，通过使用功能性的体外重构异染色质系统来更好 了解在孤立的环境中异质变沉默的机制。其次，使用pre- 组装的异染色质结构域，以识别细胞内有助于异染色质的新因素 编队和维护。第三，通过建立一个系统来解决特定非编码的贡献 RNA元素启动和维持异染色质的元素。 在异染色质的酵母模型中的初步研究将对现场做出重要贡献 染色质生物学，还将作为在人类系统中进行后续研究的模型 过渡到独立研究员之后。该项目的主要目标是全面 表征异染色质结构域，以比较与基因核心的保守特征 沉默和基因组稳定性。人类异质域的多样性很明显，但一般 管理这些领域组装的原则与萌芽和裂变酵母中的原则非常相似 型号。这些基本的相似性使得将酵母研究结果与人类进行比较以及 使用在较简单的酵母模型中开发的方法作为研究人类的指南 异染色质。这个主题是该提议的基础，以及我过渡到我将采取的方向 独立。