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

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 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，涉及数百个蛋白质因子和几个非编码的SnRNA。RNA前剪接过程需要移除被称为内含子的RNA片段，如果不移除内含子，将干扰该基因的正常表达。几乎所有的人类基因都含有内含子，而要想正常运作，就必须忠实而快速地移除这些内含子。真核细胞中负责去除内含子的机制被称为剪接体，它是一组RNA和许多蛋白质的集合，在单个内含子去除的过程中多次重排，以高度协调的方式招募因子和排出其他因子。最近在酵母细胞中发现了五个SnRNP，这对剪接体组装机制中的一个范式提出了挑战，特别是剪接体组装在很大程度上是预先组装的，重排发生在这个预先组装的复合体中。单细胞真核生物酿酒酵母可用于研究剪接体的组装和前mRNA剪接反应的过程，因为它可以进行遗传和生化分析，并以与人类剪接反应相同的化学方式执行前mRNA剪接反应。这个职业项目将解决剪接体组装的机制和五个SnRNP与其前mRNA底物结合的方式。对早期剪接中间产物的组成和功能的分析将允许对剪接体组装中的初始事件进行全面的生化和遗传分析。与奥斯汀独立学区合作，该项目还努力发起和执行一项全面的外展计划，让该地区的高中教师、管理人员和学生参与，为感兴趣的初中生和高中生提供接触现代分子生物学以及他们可供选择的教育和研究的机会。

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.