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.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 对于每个真核生物来说，一个复杂的问题是如何调节染色质重塑的过程，这是介导整个基因组DNA可及性的关键功能。使用UCSF质谱设备，我们发现RSC重塑复合物的一个重要亚单位Rsc 4在其N-末端的K25上被Gcn 5乙酰化。 在标准生长条件下纯化RSC表明大部分Rsc 4是乙酰化的。 野生型和突变型（K25 R）RSC复合物的体外比较揭示了类似的能力，重塑核小体，结合DNA，并识别乙酰化。 由于乙酰化似乎不会改变RSC的酶性质，我们正在采取几种体内方法来了解乙酰化是重要的生物过程，并了解乙酰化的功能后果。 具体来说，我们将使用质谱法来确定乙酰化被调节的条件，并结合基因突变，确定负责这种调节的基因。 这些研究是第一个表征乙酰化的染色质重塑复合物的背景下的功能。