The Role of Protein Arginine Methylation in the Co-transcriptional Recruitment of pre-mRNA Splicing Factors

蛋白质精氨酸甲基化在前体 mRNA 剪接因子共转录招募中的作用

• 批准号: 1051350
• 负责人: Michael Yu
• 金额: $ 75万
• 依托单位: SUNY at Buffalo
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1051350&HistoricalAwards=false
• 关键词: Role; Protein; Arginine; Methylation; Co

Intellectual Merit. In eukaryotes, proper gene expression is vital for the control of many cellular processes and responses. The initial step of gene expression requires the copying of the DNA into an intermediate molecule called the RNA via a process called transcription. During transcription, the unfinished RNA must simultaneously be processed by a multitude of specific proteins, including the splicing factors (which are members of a macromolecular protein complex called the spliceosome), in order to generate a final, finished product. As the RNA molecules are made and processed, they are packaged into a complex called messenger ribonucleoprotein (mRNP) particle and once finished, the entire mRNP is then exported out of the cell nucleus. To achieve and coordinate the coupling of these biological processes, the RNA-binding proteins must be recruited to the RNA in a very precise order during transcription. As a way to expand or to regulate the function of a protein, many RNA-binding proteins are modified chemically by specific enzymes. Arginine methylation is one such type of chemical modification that is found on many RNA binding proteins, and this modification is catalyzed by a family of enzymes known as protein arginine methyltransferases (PRMT). This enzyme family has been identified in species ranging from the budding yeast to humans, with the most conserved member of this family being PRMT1. In the budding yeast Saccharomyces cerevisiae, it was established that yeast PRMT1 (termed Hmt1) plays a functional role in promoting the recruitment of splicing factors during transcription, which is vital to mRNP formation. The goals of this project are to examine the mechanisms by which protein arginine methylation modulates the co-transcriptional recruitment of splicing factors in the simple eukaryotic model organism Saccharomyces cerevisiae. A combination of molecular biology, biochemistry, proteomic, and genomic methodologies will be used. Because of the high conservation between yeast Hmt1 and its homologs in higher eukaryotes, the information obtained from this project will provide insights that will likely be applicable to those organisms.Broader Impacts. This project will provide opportunities to train graduate students and post-doctoral fellows at University at Buffalo, which is a very diverse campus. The graduate students will receive training in the areas of molecular biology, biochemistry, and genomics. Undergraduate students will also be recruited to take part in various aspects of this project and these recruits will include members of the New York Collegiate Science & Technology Entry (CSTEP) program, an enrichment program for students who are economically disadvantaged or historically underrepresented. Since the project involves the use of molecular biology with genomics and computational analysis, the students will benefit from crossdisciplinary training. Travel funds will be used to allow students to present talks and posters at national scientific meetings. Findings from this work will be incorporated into an Advanced Molecular Biology laboratory course offered to the more senior undergraduates at University at Buffalo. The microarray data generated from this project will be made available to the public.

知识价值。在真核生物中，适当的基因表达对于控制许多细胞过程和反应至关重要。基因表达的第一步需要通过转录过程将DNA复制到称为RNA的中间分子中。在转录过程中，未完成的RNA必须同时被许多特定的蛋白质加工，包括剪接因子（剪接体是一种大分子蛋白质复合物的成员），以产生最终的成品。当RNA分子被制造和加工时，它们被包装成一种称为信使核糖核蛋白（mRNP）颗粒的复合物，一旦完成，整个mRNP就会被输出到细胞核外。为了实现和协调这些生物过程的耦合，RNA结合蛋白必须在转录过程中以非常精确的顺序被招募到RNA上。作为一种扩展或调节蛋白质功能的方法，许多rna结合蛋白通过特定的酶进行化学修饰。精氨酸甲基化是在许多RNA结合蛋白上发现的一种化学修饰，这种修饰是由一系列被称为蛋白质精氨酸甲基转移酶（PRMT）的酶催化的。这个酶家族已经在从芽殖酵母到人类的各种物种中被发现，这个家族中最保守的成员是PRMT1。在出芽酵母Saccharomyces cerevisiae中，已经确定酵母PRMT1（称为Hmt1）在转录过程中促进剪接因子的募集中发挥功能作用，这对mRNP的形成至关重要。本项目的目的是研究蛋白质精氨酸甲基化调节简单真核模式生物酿酒酵母剪接因子共转录募集的机制。结合分子生物学，生物化学，蛋白质组学和基因组的方法将被使用。由于酵母Hmt1及其同源物在高等真核生物中的高度保守性，从该项目中获得的信息将提供可能适用于这些生物的见解。更广泛的影响。该项目将为布法罗大学培养研究生和博士后提供机会，布法罗大学是一个非常多元化的校园。研究生将接受分子生物学、生物化学和基因组学领域的培训。本科生也将被招募来参与这个项目的各个方面，这些新兵将包括纽约大学科学与技术入门（CSTEP）计划的成员，这是一个为经济上处于不利地位或历史上代表性不足的学生提供的充实计划。由于该项目涉及到分子生物学与基因组学和计算分析的结合，学生将受益于跨学科的训练。旅行经费将用于学生在国家科学会议上发表演讲和海报。这项工作的发现将被纳入布法罗大学高级本科生的高级分子生物学实验课程。该项目产生的微阵列数据将向公众开放。