INVESTIGATING THE ROLE OF RSC4 ACETYLATION

研究 RSC4 乙酰化的作用

• 批准号: 8363797
• 负责人: GEETA J NARLIKAR
• 金额: --
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363797
• 关键词: Acetylation; Biological Process; Chromatin Remodeling Factor; Complex; DNA; DNA Binding; Funding; Gene Mutation; Genes; Genome; Grant; Growth; In Vitro; Mass Spectrum Analysis; Mediating; National Center for Research Resources; Nucleosomes; Organism; Principal Investigator; Process; Property; Regulation; Research; Research Infrastructure; Resources; Role; Source; United States National Institutes of Health; chromatin remodeling; cost; in vivo; mutant

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. A complex problem for every eukaryotic organism is how to regulate the process of chromatin remodeling, a critical function to mediate DNA accessibility throughout the genome. Using the UCSF Mass Spec Facility, we have found that Rsc4, an essential sub unit of the RSC remodeling complex, is acetylated, by Gcn5, on K25 at its N-terminus. Purification of RSC under standard growth conditions suggests that most of Rsc4 is acetylated. In vitro comparisons of WT and mutant (K25R) RSC complexes reveal similar capabilities to remodel nucleosomes, bind DNA, and recognize acetylation. Since acetylation does not appear to alter the enzymatic properties of RSC, we are taking several in vivo approaches to understand the biological processes in which acetylation is important and to understand the functional consequences of the acetylation. Specifically, we will use mass spectrometry to identify conditions where acetylation is regulated and, combined with genetic mutations, identify the genes responsible for this regulation. These studies are the first to characterize the function of acetylation in the context of a chromatin remodeling complex.

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。 每个真核生物的一个复杂问题是如何调节染色质重塑的过程，这是介导整个基因组中DNA可及性的关键功能。使用UCSF质量规格功能，我们发现RSC4是RSC重塑复合物的必不可少的子单元，由GCN5在其N末端的K25上被GCN5乙酰化。 在标准生长条件下RSC的纯化表明RSC4的大多数都是乙酰化的。 WT和突变体（K25R）RSC络合物的体外比较揭示了与重塑核小体，结合DNA和识别乙酰化的相似能力。 由于乙酰化似乎并没有改变RSC的酶促特性，因此我们采用了几种体内方法来了解乙酰化很重要并了解乙酰化的功能后果的生物学过程。 具体而言，我们将使用质谱法来确定调节乙酰化的条件，并与遗传突变结合确定负责该调节的基因。 这些研究是第一个在染色质重塑络合物的背景下表征乙酰化功能的研究。