STRUCTURE DETERMINATION OF THE DNA BINDING DOMAIN OF S CEREVISIAE CHD1 IN COMPL

完整的酿酒酵母CHD1 DNA结合域的结构测定

• 批准号: 8363342
• 负责人: GREGORY DEAN BOWMAN
• 金额: $ 0.62万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363342
• 关键词: ATP phosphohydrolase; Binding; C-terminal; Chromatin; Chromatin Structure; Communication; Complex; Crystallography; DNA; DNA Binding; DNA Binding Domain; DNA Repair; Enzymes; Eukaryotic Cell; Event; Funding; Genetic Transcription; Grant; Light; Maps; Modeling; Mono-S; Motor; N-terminal; National Center for Research Resources; Nucleosomes; Nucleotides; Oligonucleotides; Physiological; Principal Investigator; Recruitment Activity; Research; Research Infrastructure; Resources; Saccharomyces cerevisiae; Site; Source; Stretching; Structure; Synchrotrons; United States National Institutes of Health; chromatin remodeling; cost; helicase; insight; promoter

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Chromatin organization pertains to the packaging of long stretches of DNA in eukaryotic cells, with the basic unit of such structure being mono-nucleosomes wrapped around by ~146bp of DNA. This structural organization of DNA leads to leads to the occlusion of functionally important sites (such as promoters), thereby hindering transcription and other crucial physiological events such as replication and DNA repair. In order to facilitate transcription and other DNA-related transactions, enzymes called chromatin-remodelers are recruited to nucleosomes. However, the step-by-step mechanism by which remodeling of chromatin structure occurs is yet to be unraveled. We seek to understand the mechanism of chromatin remodeling using Chd1, a monomeric chromatin remodeler with an N-terminal double chromodomain, a central Helicase-like ATPase motor and a C-terminal DNA binding domain. The DNA binding domain is an ~275 residues stretch with a myb-homology region and has been shown to interact with DNA in a sequence non-specific manner. Additionally, studies have shown that the DNA-binding domain is vital for efficient remodeling by Chd1. In order to understand the manner in which the DNA-binding module of S.cerevisiae Chd1 interacts with DNA, we have crystallized the DNA binding domain in complex with several oligo-nucleotides and seek to determine the structure of the complex. Elucidation of the crystal structure of this complex would(i) help ascertain if this domain aids the distortion of DNA upon binding to it, (ii) facilitate mapping of residues important for interaction with nucleosomal DNA, (iii) restrict models for nucleosome-remodeler complexes, and (iv) provide insight into domain-domain communication in Chd1.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 染色质组织涉及真核细胞中长段 DNA 的包装，这种结构的基本单位是由约 146bp 的 DNA 包裹的单核小体。 DNA 的这种结构组织导致功能重要位点（例如启动子）的闭塞，从而阻碍转录和其他重要的生理事件，例如复制和 DNA 修复。为了促进转录和其他 DNA 相关交易，称为染色质重塑酶的酶被招募到核小体中。 然而，染色质结构重塑发生的逐步机制尚未阐明。我们试图了解使用 Chd1 进行染色质重塑的机制，Chd1 是一种单体染色质重塑剂，具有 N 端双染色质结构域、中央解旋酶样 ATP 酶马达和 C 端 DNA 结合域。 DNA 结合域是一段约 275 个残基的片段，具有 myb 同源区域，并已被证明以序列非特异性方式与 DNA 相互作用。此外，研究表明 DNA 结合域对于 Chd1 的有效重塑至关重要。为了了解酿酒酵母 Chd1 的 DNA 结合模块与 DNA 相互作用的方式，我们将 DNA 结合域与几种寡核苷酸形成复合物结晶，并试图确定该复合物的结构。阐明该复合物的晶体结构将(i)帮助确定该结构域是否有助于DNA在与其结合时变形，(ii)促进对与核小体DNA相互作用重要的残基的映射，(iii)限制核小体-重塑复合物的模型，以及(iv)提供对Chd1中结构域-结构域通信的深入了解。