Metal Ion-dependent Folding of the Spliceosomal U2-U6 snRNA Complex

该项目的目标是了解剪接体的RNA和蛋白质组分如何组装以执行剪接化学。剪接体是负责从前体信使（pre-m）RNA分子中去除非编码区域的细胞装置。单个RNA组分的折叠以及RNA-RNA相互作用形成的结构的折叠是剪接体组装的一个重要特征。剪接体的小核（sn）RNA组分对金属离子的特异性结合对于组装和催化活性是必不可少的，但对单个结合位点或金属离子折叠的作用知之甚少。发光和溶液核磁共振技术将用于研究由成对的U2和U6 snrna形成的复合物中金属离子依赖的结构变化。U2和U6 snRNA是两个剪接步骤都需要的两个snRNA，它们的序列在系统发育上是高度保守的。这项工作的广泛影响包括在实验室环境中对研究生和本科生进行教育，为这些学生提供分子生物学、生物化学、结构生物学和光谱技术方面的跨学科培训；作为一名课堂教师，为学院的教学做出了贡献，特别是在核酸结构与功能研究型课程的开发和教学方面；社区外展项目，包括社区科学展览的评审；参观规模较小的本科学院，包括女子学院和历史上的黑人学校，举办研究研讨会，鼓励学生考虑攻读科学研究生课程；鼓励女性和少数族裔学生从事科学事业。后一个领域的形式是通过佛罗里达州立大学的女性数学、科学和工程项目持续指导女学生，招募少数族裔本科生到实验室和部门进行暑期研究实习，资助少数族裔学生和博士后研究员在PI的实验室做研究。

The goal of this project is to understand how RNA and protein components of the spliceosome, the cellular apparatus responsible for the removal of noncoding regions from precursor messenger (pre-m)RNA molecules, assemble to perform splicing chemistry. The folding of individual RNA components, and of structures formed by RNA-RNA interactions, is an important feature of spliceosome assembly. Specific binding of metal ions by the small nuclear (sn)RNA components of the spliceosome is essential for assembly and catalytic activity, yet little is known about individual binding sites or the role of metal ions folding. Luminescence and solution NMR techniques will be used to investigate the metal ion-dependent structural change in the complex formed by paired U2 and U6 snRNAs. U2 and U6 snRNA are the only two snRNAs required for both steps of splicing, and their sequences are highly conserved phylogenetically. Broader impacts of this work include education of graduate and undergraduate students in a laboratory setting, which provides these students with interdisciplinary training in molecular biology, biochemistry, structural biology, and spectroscopic techniques; contributions to the PI's strength as a classroom teacher, in particular development and teaching of a research-based course in Nucleic Acid Structure & Function; community outreach programs, including judging in community science fairs; visits to small undergraduate colleges, including women's and historically black schools, to present research seminars and encourage students to consider graduate programs in science; and encouragement of women and minority students to pursue careers in science. This latter area takes the form of continuous mentoring of women students through Florida State University's Women in Mathematics, Science, and Engineering Program, recruiting minority undergraduates into the lab and department for Summer Research Internships, sponsoring minority students and postdoctoral fellows for fellowships to do research in the PI's laboratory.