Structure-function relationships of metal ion-binding in the spliceosomal U2-U6 snRNA complex

该奖项是根据2009年美国复苏和再投资法案（公法111-5）资助的。智力优势：该项目的长期目标是实现对功能性RNA（核糖核酸）分子如何执行其生物活动以及金属离子在这些过程中的作用的更基本的理解。DNA（脱氧核糖核酸）的唯一生物学作用是编码细胞的遗传信息，而其化学表亲RNA的生化特性使其能够在基因表达的许多步骤中发挥关键作用，包括调节和催化。RNA能够进行广泛活动的能力既源于化学差异（具体来说，DNA的糖成分中缺少一个氧原子），也源于单链RNA折叠成无数构象的能力，每种构象都具有特定的功能。功能性RNA分子的另一个特征是能够在特定位置结合某些金属离子，从而扩展折叠RNA的结构和催化特性。本研究的目的是研究小核（sn）RNA分子的金属离子结合特性，这些分子与许多蛋白质一起形成剪接体。真核剪接体介导一种关键的活性，在前体信使RNA （pre-m）分子翻译成蛋白质之前，将非编码区从前体信使RNA分子中移除。snrna与金属离子的特异性相互作用对于剪接体的组装和催化活性至关重要，但对单个结合位点的详细结构以及这些金属离子在催化活性中的作用知之甚少。这项研究的意义在于它有可能更详细地了解rna -金属离子配合物的结构和化学性质之间的相互关系。这些数据将极大地增强对rna介导的催化过程的基本理解，并将产生新的方法。这项工作的更广泛的影响包括：1)在实验室教育研究生和本科生，为他们提供分子生物化学、RNA结构生物学和光谱技术的跨学科培训；2)作为一名课堂教师，为学院的实力做出了贡献（2004年获本科教学奖），并扩展了学院开发的“核酸结构与功能”研究型课程；3)招募访问小型文理本科学院，目标是女性和历史上的黑人学校；4)鼓励妇女和少数民族学生从事科学事业。后一个领域采取的形式是将少数民族本科生带到实验室和部门进行暑期研究实习，资助少数民族学生和博士后在她的实验室做研究。PI最近将她的实验室从佛罗里达州立大学搬到了纽约城市大学（CUNY）的亨特学院，在那里她的团队将可以使用优秀的研究设施，特别是在一个完全翻新的核磁共振设施中使用新的核磁共振（NMR）光谱仪。亨特是一所少数族裔服务机构，65%的女性学生毕业于全国5%的物理科学博士学位。该提案由基因和基因组系统集群和生物分子系统集群共同资助。

This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5). Intellectual Merit: The long-range objective of this project is to achieve a more fundamental understanding of how functional RNA (ribonucleic acid) molecules perform their biological activities, and the role of metal ions in these processes. Whereas the sole biological role of DNA (deoxyribonucleic acid) is to encode a cell's genetic information, the biochemical properties of RNA, its chemical cousin, enable it to mediate critical roles in numerous steps in gene expression, including regulation and catalysis. The power of RNA to perform its wide range of activities stems both from chemical differences (specifically, an oxygen atom missing from the sugar component of DNA) and from the ability of single-stranded RNA to fold into myriad conformations, each capable of a specific function. An additional feature of functional RNA molecules is the ability to bind certain metal ions in specific locations, which extend the structural and catalytic properties of the folded RNA. The aims of this research are to examine the metal ion binding properties of the small nuclear (sn)RNA molecules that, together with numerous proteins, form the spliceosome. The eukaryotic spliceosome mediates a pivotal activity that removes noncoding regions from precursor messenger (pre-m)RNA molecules prior to their translation into protein. Specific interaction with metal ions by snRNAs is essential for spliceosome assembly and catalytic activity, yet little is known about the detailed structure of individual binding sites and the role of these metal ions in catalytic activity. The significance of this research lies in its potential to achieve a more detailed understanding of the interrelationship between structural and chemical properties of RNA-metal ion complexes. These data will greatly enhance fundamental understanding of the processes involved in RNA-mediated catalysis and will generate new methods. The broader impact of this work includes: 1) education of graduate and undergraduate students in a laboratory that provides them with interdisciplinary training in molecular biochemistry, RNA structural biology, and spectroscopic techniques; 2) contributions to the PI's strength as a classroom teacher (for which she won an undergraduate Teaching Award in 2004), and expansion of a research-based course in Nucleic Acid Structure & Function developed by the PI; 3) recruiting visits to small liberal arts undergraduate colleges, targeting women's and historically black schools; 4) encouragement of women and minority students to pursue careers in science. This latter area takes the form of bringing minority undergraduates into the laboratory and department for summer research internships, sponsoring minority students and postdoctoral fellows for fellowships to do research in her laboratory. The PI has recently moved her laboratory from Florida State University to Hunter College of the City University of New York (CUNY), where her group will have access to outstanding research facilities, specifically a new nuclear magnetic resonance (NMR) spectrometer in a completely renovated NMR facility. Hunter is a minority serving institution with 65% women students that graduates 5% of the nation's Ph.D.s in the physical sciences. This proposal is being co-funded by the Genes & Genomes Systems Cluster and the Biomolecular Systems Cluster.