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.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 染色质组织与真核细胞中长长的DNA的包装有关，这种结构的基本单位是单核小体的DNA包裹的单核小体。 DNA的结构组织导致导致功能重要的位点（例如启动子）的阻塞，从而阻碍转录和其他关键的生理事件，例如复制和DNA修复。为了促进转录和其他与DNA相关的交易，将称为染色质复制器的酶募集到核小体中。 但是，尚未阐明染色质结构重塑的分步机制。我们试图使用CHD1（一种具有N末端双染色体蛋白酶，中央解旋酶样ATPase运动和C-末端DNA结合结构蛋白的单体染色质重塑剂CHD1）来理解染色质重塑的机制。 DNA结合结构域是一个〜275个残基，具有MYB - 同学区域，并已显示出以非特异性方式与DNA相互作用。此外，研究表明，DNA结合域对于通过CHD1有效重塑至关重要。为了理解Cerevisiae Chd1的DNA结合模块与DNA相互作用的方式，我们已经结晶了与几种寡核苷酸的复合物中的DNA结合结构域，并寻求确定复合物的结构。阐明该复合物的晶体结构将有助于确定该结构域在与其结合时是否有助于DNA的变形，（ii）促进对与核小体DNA相互作用重要的残基的映射，（iii）限制了核小体 - 变形剂的模型复合物和（iv）提供了对CHD1中域域通信的见解。