CAREER: MECHANISM OF PENTA-SNRNP FUNCTION IN PRE-MRNA SPLICING

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 基因在我们所有细胞中的表达方式是一个高度调控的复杂过程，涉及数百种蛋白质因子和几种非编码snRNAs。前mRNA剪接的过程需要去除称为内含子的RNA片段，如果不去除内含子，就会干扰该基因的正常表达。几乎所有人类基因都含有内含子，需要忠实而快速地去除它们才能正常发挥作用。真核细胞中负责去除内含子的机制被称为剪接体，它是RNA和许多蛋白质的集合，在单个内含子去除过程中会进行多次重排，以高度协调的方式招募因子并排出其他因子。最近在酵母细胞中发现的penta-snRNP挑战了剪接体组装机制中的范式，具体地说，该机制主要是预组装的，并且重排发生在该预组装的复合物中。单细胞真核生物酿酒酵母将用于研究剪接体组装和前mRNA剪接反应的过程，因为它适合遗传和生化分析，并以与人类剪接反应化学相同的方式执行前mRNA剪接反应。这个CAREER项目将解决剪接体组装的机制和penta-snRNP接合其前体mRNA底物的方法。早期剪接中间体的组成和功能分析将允许在剪接体组装的初始事件进行全面的生化和遗传分析。与奥斯汀独立学区一起，该项目还致力于发起和执行一个全面的外展计划，涉及地区高中教师，管理人员和学生，为感兴趣的初中和高中学生提供现代分子生物学和教育和研究选择。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The way in which genes are expressed in all of our cells is a highly regulated, complex process involving hundreds of protein factors and several non-coding snRNAs. The process of pre-mRNA splicing is required to remove stretches of RNA called introns, which if not removed would interfere with proper expression of that gene. Almost all human genes contain introns and their faithful and rapid removal is required for proper functioning. The machinery in eukaryotic cells charged with the removal of introns is called the spliceosome, a collection of RNAs and many proteins that rearranges several times during the course of a single intron removal, recruiting factors and ejecting others in a highly coordinated manner. The recent discovery of the penta-snRNP in yeast cells has challenged a paradigm in the mechanism of spliceosome assembly, specifically, that the machinery is largely pre-assembled and that the rearrangements occur within this pre-assembled complex. The single-celled eukaryotic organism Saccharomyces cerevisiae will be used to study the process of spliceosome assembly and the pre-mRNA splicing reaction as it is amenable to genetic and biochemical analysis and executes the pre-mRNA splicing reaction in a way that is chemically identical to the human splicing reaction. This CAREER project will address the mechanism of spliceosome assembly and the means by which the penta-snRNP engages its pre-mRNA substrate. Compositional and functional analyses of the early splicing intermediates will allow a comprehensive biochemical and genetic analysis of the initial events in spliceosome assembly. In conjunction with the Austin Independent School District, this project also endeavors to initiate and execute a comprehensive outreach program involving area high school teachers, administrators and students to provide exposure of interested junior and senior students to modern molecular biology and the educational and research options available to them.