Structural biology of chromatin in vitro and in cells

体外和细胞内染色质的结构生物学

基本信息

批准号：
10797358
负责人：
Galia Debelouchina
金额：
$ 2.64万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2025-06-30
项目状态：
未结题

项目摘要

Project Summary The laboratory led by PI Galia Debelouchina has the following objectives: 1) The development of structural biology methodology to study complex and dynamic biological assemblies in vitro, and 2) The extension of these methodologies for structural biology investigations in the cellular environment. Our methodology development combines solid-state nuclear magnetic resonance (NMR) spectroscopy with state-of-the-art chemical biology tools for the comprehensive description of biological systems both in vitro and in cells. We use these technologies to investigate chromatin, a complex protein-DNA polymer responsible for the packaging of genetic information in the nucleus. Over the next five years, we plan to focus on the following projects: 1) The influence of protein post-translational modifications on chromatin structure and dynamics. Modifications such as methylation and acetylation have profound effects on the compaction of the chromatin polymer and the accessibility of the packaged DNA. We plan to investigate the molecular mechanisms of chromatin compaction and decompaction imparted by these modifications at atomic resolution using solid-state NMR spectroscopy. We expect that this study will provide unique insights into the biophysical properties of the chromatin polymer and how they can be translated into functional biological outputs. 2) The molecular basis of heterochromatin formation. Heterochromatin compartments are associated with gene silencing and repetitive DNA sequences, and their formation is a vital step in cell differentiation and development. Recent hypotheses indicate that they are formed through liquid-liquid phase separation. We plan to investigate the interactions that promote phase separation and to provide the first molecular description of this process. 3) Development of NMR-based tools for structural biology in cells. Here, we plan to focus on the design and implementation of a strategy to target a specific protein in the cellular milieu and to increase its NMR signals selectively over the cellular background. Ultimately, we aim to develop a selective and sensitive NMR methodology that will allow us to record relevant structural data of endogenous amounts of cellular proteins. Such tools will provide the unprecedented opportunity to build an atomic resolution picture of the cellular milieu and to investigate changes in protein structure and dynamics as we stress, age or treat cells with therapeutic agents.

项目摘要 Pi Galia Debelouchina领导的实验室具有以下目标：1）结构的发展在体外研究复杂和动态生物组装的生物学方法论，2）这些用于细胞环境中结构生物学研究的方法。我们的方法论开发结合了固态核磁共振（NMR）光谱与最先进的光谱化学生物学工具，用于体外和细胞中生物系统的全面描述。我们使用这些技术研究染色质，这是一种复杂的蛋白-DNA聚合物核中遗传信息的包装。在接下来的五年中，我们计划专注于以下项目：1）翻译后修饰对染色质结构和动力学的影响。修饰（例如甲基化和乙酰化）对染色质的压实具有深远的影响聚合物和包装DNA的可访问性。我们计划研究这些修饰在原子分辨率下使用固态赋予这些修饰的染色质压实和分解 NMR光谱。我们预计这项研究将为您的生物物理特性提供独特的见解染色质聚合物及其如何转化为功能性生物产量。 2）分子基础异染色质形成。异染色质区室与基因沉默和重复有关 DNA序列及其形成是细胞分化和发育中的重要步骤。最近的假设表明它们是通过液体液相分离形成的。我们计划调查互动促进相位分离并提供该过程的第一个分子描述。 3）开发基于NMR的细胞结构生物学工具。在这里，我们计划专注于设计和实施靶向细胞环境中特定蛋白质并选择性增加其NMR信号的策略细胞背景。最终，我们旨在开发一种选择性和敏感的NMR方法，该方法将允许美国记录内源量的细胞蛋白的相关结构数据。这样的工具将提供前所未有的机会来建立蜂窝环境的原子分辨率图片并调查当我们用治疗剂压力，年龄或治疗细胞时，蛋白质结构和动态的变化。