Structural and Functional Analysis of the CHD1 Chromatin Remodeler

CHD1 染色质重塑蛋白的结构和功能分析

• 批准号: 8043586
• 负责人: GREGORY DEAN BOWMAN
• 金额: $ 27.91万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8043586
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; Architecture; Binding; Biochemical; C-terminal; Cells; Chromatin; Chromatin Remodeling Factor; Chromatin Structure; Chromosome Structures; Chromosomes; Communication; Complex; Congenital Abnormality; Coupling; DNA; DNA Binding; DNA Footprint; Disease; Docking; Elements; Enzymes; Future; Gene Expression; Gene Expression Regulation; Gene Silencing; Genes; Genome; Goals; Health; Histones; Human body; Hydrolysis; Individual; Inherited; Knowledge; Link; Malignant Neoplasms; Measurement; Modeling; Modification; Molecular; Molecular Profiling; Motion; Movement; N-terminal; Nucleosomes; Pattern; Play; Positioning Attribute; Process; Proteins; Reaction; Relative (related person); Resolution; Roentgen Rays; Role; Site-Directed Mutagenesis; Slide; Staging; Structure; Surface; Techniques; Therapeutic; Tissues; Variant; Work; X-Ray Crystallography; chromatin remodeling; combat; design; developmental disease; genome-wide; human disease

DESCRIPTION (provided by applicant): The physical organization of chromosomes is intimately tied to gene regulation. Chromatin remodeling refers to the dynamic compaction and decondensation of eukaryotic chromosomes through the covalent modification and physical movement of nucleosomes. Knowledge of the processes that drive remodeling are essential for gaining a more insightful understanding of human diseases that result from disruptions of normal gene regulation, such as cancer and inherited developmental disorders. Relatively little is known regarding the mechanism by which ATP-dependent remodeling factors alter nucleosome structure, with no understanding of how distinct remodeler domains functionally interact to promote remodeling, how key remodeler domains are positioned in space with respect to one another, nor how remodelers embrace the nucleosome substrate. The long-term objective of this proposal is to establish a biochemical and biophysical framework necessary for describing and understanding the process of chromatin remodeling. This proposal focuses on the CHD1 chromatin remodeling factor, with specific aims to (1) dissect the CHD1 remodeling cycle by characterizing partially dysfunctional variants, (2) determine the architecture of the CHD1 remodeler using X-ray crystallography, and (3) determine the organization of a CHD1:nucleosome complex using small angle X-ray scattering (SAXS) and DNA footprinting. By coupling functional studies of CHD1 with structural information gained through X-ray crystallography, SAXS, and DNA footprinting, we expect to elucidate key steps of the remodeling reaction. In the future, a deeper understanding of chromatin remodeling will be essential for developing therapeutics that manipulate genome-wide gene expression for treatment of human disease. PUBLIC HEALTH RELEVANCE Nearly all cells in the human body possess the same set of genes, yet only a subset of these genes are turned "on" in any particular tissue. The "on" and "off" states of genes are regulated in a complex and ill-defined manner that directly correlates with the physical packaging of chromosomes, called the chromatin structure. Understanding the process by which chromosomes can be unpackaged and repackaged by so-called chromatin remodeling factors is necessary for understanding and therefore combatting many diseases such as cancer where there are imbalances in gene expression.

描述（由申请人提供）：染色体的物理组织与基因调节密切相关。染色质重塑是指通过核小体的共价修饰和物理运动对真核染色体的动态压实和反应。了解驱动重塑的过程对于获得对正常基因调节（例如癌症和遗传性发育障碍）中断的人类疾病的更深入的理解至关重要。关于ATP依赖性重塑因子改变核小体结构的机制，相对鲜为人知，没有了解独特的重塑域在功能上如何相互作用以促进重塑，关键重塑域如何相对于彼此的空间位置，也不是重塑剂如何拥抱重塑剂。核小体底物。该建议的长期目标是建立一个生化和生物物理物理框架，用于描述和理解染色质重塑的过程。该提案的重点是CHD1染色质重塑因子，具体目的是通过表征部分功能障碍的变体来解剖CHD1重塑周期，（2）确定使用X射线晶体术的CHD1重塑剂的结构，并确定（3）确定（3）确定（3）使用小角度X射线散射（SAXS）和DNA足迹的CHD1：核小体复合物的组织。通过将CHD1的功能研究与通过X射线晶体学，SAXS和DNA足迹获得的结构信息耦合，我们希望阐明重塑反应的关键步骤。将来，对染色质重塑的更深入了解对于开发操纵全基因组基因表达以治疗人类疾病的治疗剂至关重要。公共卫生相关性几乎所有人体中的所有细胞都具有相同的基因，但是在任何特定组织中，只有一部分这些基因被打开。基因的“ ON”和“ OFF”状态以复杂且不确定的方式调节，该方式与称为染色质结构的染色体的物理包装直接相关。了解可以通过所谓的染色质重塑因子解开和重新包装染色体的过程，对于理解，因此与许多基因表达失衡的癌症等许多疾病作斗争。