RNA PSEUDOKNOT STRUCTURE AND STABILITY

我的长期职业目标是保持一个强大的研究计划， 具有重要的生物医学意义。 的头几年 作为一名独立的科学家，我相信我已经做出了一个 对我们理解RNA结构的重大贡献， RNA-蛋白质识别和结合的结构基础 伪结，以及与之相关的平移装置的组成部分 核糖体移码和重新编码。提供的工资支持 由独立科学家奖（K 02）将减轻我的一半， 目前的教学量（目前每年约80次讲座）， 并提供行政职责的减免， 我的研究时间。 提供的额外研究时间 这项独立科学家奖将立即产生积极的影响， 对我的NIH资助的研究生产力的影响。 目前的提案是基于我最近资助的NIH（R 01）提案， 旨在理解结构之间的关系， 稳定性和功能的一个家庭的相关RNA假结。 假结是RNA结构的一个元素，广泛存在于 自然，在各种生物过程中发挥着不可或缺的作用。 研究将集中在来自基因32 mRNA的RNA假结， 噬菌体T2和SRV-1的gag-pro mRNA移码位点 慢病毒初步证据显示， 噬菌体和这些假结是一种 一个大家族的相关RNA假结，其成员 功能不同，但有共同的结构基序。 以下 实验结果表明：（1）详细的结构， 噬菌体T2和SRV-1假结将使用多克隆抗体测定。 三维NMR方法。 伪结结构的比较将 提供了关于假结的共同特征 由于功能和结构的原因，家族是保守的。 （二） 以前的结果表明，SRV-1移码信号也可能是 嵌入伪结的稳定性、动力学和展开 路径，以及结构。 一系列核磁共振实验将在 用于详细研究RNA的这些动态方面， 伪结

My long-term career goal is to maintain a strong research program in an area of fundamental biomedical importance. In the first few years of my career as an independent scientist, I believe I have already made a substantial contribution to our understanding of RNA structure, the structural basis of RNA-protein recognition and binding, RNA pseudoknots, and components of the translational apparatus associated with ribosomal frameshifting and re-coding. The salary support provided by an Independent Scientist Award (K02) will relieve me of half of my present teaching load (currently approximately 80 lectures per year), and provide relief from administrative duties, freeing 75 to 80 percent of my time for research. The additional research time made available by this Independent Scientist Award will have an immediate and positive effect on my NIH-funded research productivity. The present proposal is based on my recently funded NIH (R01) proposal, directed toward understanding the relationships between structure, stability and function in a family of related RNA pseudoknots. Pseudoknots are an element of RNA structure that is widespread in nature, playing an integral role in a variety of biological processes. Investigations will focus on RNA pseudoknots from the gene 32 mRNA of bacteriophage T2, and the gag-pro mRNA frameshift site of the SRV-1 lentivirus. Preliminary evidence strongly suggests that the bacteriophage and these pseudoknots are representative members of a single large family of related RNA pseudoknots, the members of which differ in function, but share a common structural motif. The following experiments are proposed: (1) The detailed structures of the bacteriophage T2 and SRV-1 pseudoknots will be determined using multi- dimensional NMR methods. A comparison of the pseudoknot structures will provide insight as to which of the common features of the pseudoknot family are conserved for functional versus structural reasons. (2) Previous results suggest that the SRV-1 frameshift signal may also be embedded within the pseudoknot stability, dynamics and unfolding pathway, as well as the structure. A series of NMR experiments will be used to investigate in detail these dynamic aspects of the RNA pseudoknots.