Discovering new writers and patterns underlying the CTD code

RNA聚合酶II(POL II)是将真核细胞中的大部分基因转录成RNA的蛋白质机器。POL II核心成分由12种蛋白质组成，最大蛋白质的末端称为C-末端结构域(CTD)，是与其他蛋白质和细胞机相互作用的部位。在这个项目中，我们将学习面包酵母细胞中CTD的化学变化是如何影响这些相互作用的。由于CTD相关机制在进化中是高度保守的，这项研究将使我们能够理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来扩大学生和同事的视野，这些团队带来来自多个学科的专业知识，培训下一代参与全球事务的科学家/技术专家，并为科学界提供广泛的使能工具。为了让下一代学生以及更大的社区参与到STEM学科中来，我们将特别开发一款电脑游戏，它从收集和组装分子生物学中心法则下的基本生物学过程开始，并引导玩家识别形成“最小生命基因组”所需的特征。“CTD密码”假说假设，相关蛋白质与POL II的接触是通过顺序的CTD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出现了一个重复的七肽，它在所有五个含羟基的氨基侧链上都被修饰了。这个项目的重点是这个七肽重复序列第四位的苏氨酸的磷酸化；这种磷酸化已经被认为是指导一组特定的POL II转录的基因的转录后修饰。该项目将整合化学、生物和计算方法，以确定从酿酒酵母中磷酸化Pol II CTD中的四个苏氨酸的细胞激酶。下一阶段的实验和分析将集中于获得对该残基的磷酸化如何有助于全球与基因类特定转录控制的机械性见解。

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.