YEAST RDNA PROJECT

酵母RDNA项目

• 批准号: 8361514
• 负责人: ERICA Y JACOBS
• 金额: $ 1.3万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361514
• 关键词: Behavior; Binding Proteins; Cells; Chromatin; Funding; Gene Expression; Gene Silencing; Genes; Goals; Grant; Growth; Location; Maps; Mass Spectrum Analysis; Modeling; Modification; National Center for Research Resources; Principal Investigator; Process; Protein Biosynthesis; Proteins; Research; Research Infrastructure; Resources; Ribosomal DNA; Ribosomes; Role; Saccharomycetales; Source; Time; United States National Institutes of Health; Yeasts; chromatin protein; cost; histone modification; macromolecule

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. In order to study the role of chromatin proteins in gene expression and silencing, we are analyzing chromatin proteins at the ribosomal DNA (rDNA) locus in budding yeast. Ribosomes are required for the fundamental process of protein synthesis, and their expression is regulated according to how much proteins cells need. This in turn depends on how aggressively they are dividing. In wild type yeast growing exponentially, only about half the ~150 tandemly repeated rDNA genes are active at any time, but as growth plateaus all of the repeats become silenced. Since the repeats have the same sequence, differences in behavior between them must be caused by differences in associated factors. This project has three specific aims. First we will purify native rDNA chromatin from yeast cells. Second we will identify the bound proteins, map their locations along the rDNA, and determine how they are modified using mass spectrometry. Third we will determine differences in chromatin composition and modifications between rDNA that is ?on? (transcribed) and rDNA that is off (silent), and describe how their distributions change as the locus is progressively activated or silenced. The ultimate goal is not only a map of active and silenced rDNA chromatin (including histone modifications), but also a dynamic model of rDNA activation and silencing.

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， 包括其他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。 为了研究染色质蛋白在基因表达和沉默中的作用，我们分析了芽殖酵母核糖体DNA（rDNA）位点的染色质蛋白。核糖体是蛋白质合成的基本过程所必需的，其表达根据细胞需要多少蛋白质来调节。这反过来又取决于他们分裂的力度。在指数增长的野生型酵母中，在任何时候只有大约一半的~150个串联重复的rDNA基因是活跃的，但是随着生长平台期，所有的重复序列都变得沉默。由于重复序列具有相同的序列，它们之间的行为差异必然是由相关因素的差异引起的。 该项目有三个具体目标。首先，我们将从酵母细胞中纯化天然rDNA染色质。其次，我们将鉴定结合的蛋白质，绘制它们沿rDNA沿着的位置，并使用质谱法确定它们是如何被修饰的。第三，我们将确定染色质组成和rDNA之间的修饰差异，这是？上？（转录）和rDNA关闭（沉默），并描述它们的分布如何随着基因座逐渐激活或沉默而变化。最终的目标不仅是一个活跃的和沉默的rDNA染色质（包括组蛋白修饰）的地图，但也rDNA激活和沉默的动态模型。