Discovering new writers and patterns underlying the CTD code

发现 CTD 代码背后的新作者和模式

• 批准号: 1413547
• 负责人: Aseem Ansari
• 金额: $ 60万
• 依托单位: University of Wisconsin-Madison
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1413547&HistoricalAwards=false
• 关键词: Discovering; new; writers; patterns; underlying

RNA polymerase II (Pol II) is the protein machine that transcribes the majority of genes in the eukaryotic cell into RNA. Pol II core component consists of 12 proteins, and the tail end of the largest protein, called the C-terminal domain (CTD), is the site of interaction with other proteins and cellular machines. In this project, we will earn how these interactions are affected by chemical changes of the CTD in the baker's yeast cells. Since CTD-related mechanisms are highly conserved in evolution, the research will enable our understanding of gene expression in other eukaryotes, including humans. The program will expand the horizons of students and colleagues by working in teams that bring expertise from multiple disciplines, train the next generation of globally engaged scientists/technologists, and provide widely enabling tools to the scientific community. To engage the next generation of students in STEM subjects, as well as the larger community, we will in particular develop a computer game that begins with the collection and assembly of basic biological processes that underlie The Central Dogma of Molecular Biology and leads the players to identify features required to form a "minimal living genome."The "CTD code" hypothesis posits that the engagement of relevant proteins with Pol II is coordinated by sequential post-translational modifications of the CTD. An important feature of this unusual domain is the occurrence of a repeating hepta-peptide that is modified on all five of the hydroxyl-bearing amino side chains. This project focuses on phosphorylation of threonine in position four of this hepta-peptide repeat; such phosphorylation has been implicated in directing the post-transcriptional modification of a specific set of Pol II-transcribed genes. The project will integrate chemical, biological and computational approaches to identify cellular kinases that phosphorylate threonine four of the Pol II CTD from Saccharomyces cerevisiae. The next level of experimentation and analysis will focus on obtaining mechanistic insights into how phosphorylation of this residue contributes to global versus gene class-specific transcriptional control.

RNA聚合酶II （RNA polymerase II, Pol II）是将真核细胞中大部分基因转录成RNA的蛋白质机器。Pol II核心成分由12个蛋白组成，其中最大的蛋白的尾部称为c -末端结构域（C-terminal domain， CTD），是与其他蛋白和细胞机器相互作用的部位。在这个项目中，我们将了解这些相互作用如何受到面包师酵母细胞中CTD的化学变化的影响。由于ctd相关机制在进化过程中高度保守，该研究将使我们了解包括人类在内的其他真核生物的基因表达。该计划将通过团队合作，带来来自多个学科的专业知识，培养下一代全球参与的科学家/技术人员，并为科学界提供广泛的支持工具，从而扩大学生和同事的视野。为了吸引STEM学科的下一代学生以及更大的社区，我们将特别开发一款电脑游戏，该游戏从分子生物学中心法则基础的基本生物过程的收集和组装开始，并引导玩家识别形成“最小活基因组”所需的特征。“CTD编码”假说认为，相关蛋白与Pol II的结合是通过CTD的顺序翻译后修饰来协调的。这个不寻常结构域的一个重要特征是在所有五个含羟基氨基侧链上都修饰了一个重复的七肽。这个项目的重点是在这个七肽重复的位置4的苏氨酸磷酸化；这种磷酸化与指导一组特定的Pol ii转录基因的转录后修饰有关。该项目将整合化学、生物学和计算方法，以鉴定使酿酒酵母Pol II CTD中的苏氨酸4磷酸化的细胞激酶。下一阶段的实验和分析将集中于获得关于该残基的磷酸化如何促进全局与基因类特异性转录控制的机制见解。