RNA PSEUDOKNOT STRUCTURE AND STABILITY

我的长期职业目标是保持一个强大的研究项目 具有基本生物医学重要性的领域。在…的头几年 我作为一名独立科学家的生涯，我相信我已经做出了 对我们理解RNA结构的重大贡献， 蛋白质识别和结合的结构基础 伪结点和相关联的平移设备的部件 通过核糖体的移码和重新编码。提供的工资支持 独立科学家奖(K02)将使我失去一半的 目前的教学负担(目前每年约80场讲座)， 并提供行政职责的减免，将75%至80%的 我的研究时间。提供的额外研究时间 由这一独立科学家奖将产生立竿见影的积极作用 对我的NIH资助的研究效率的影响。 目前的提案是基于我最近资助的NIH(R01)提案， 旨在理解结构之间的关系， 一个相关的RNA伪结点家族的稳定性和功能。 伪结点是RNA结构中广泛存在的一种元素 自然，在各种生物过程中发挥着不可或缺的作用。 调查将集中在来自32mRNAs基因的RNA伪结点 噬菌体T2和SRV-1的gag-promRNA移码位点 慢病毒。初步证据有力地表明， 噬菌体和这些伪结是具有代表性的成员 单个相关的RNA伪结点大家族，其成员 功能不同，但有一个共同的结构主题。以下是 提出了以下实验建议：(1)详细的结构 噬菌体T2和SRV-1伪结点将用多个 三维核磁共振方法。对伪结结构的比较将 提供有关伪结的哪些常见特征的洞察 出于功能上的原因而不是结构上的原因，家庭是保守的。(2) 先前的结果表明，SRV-1的移码信号也可能是 嵌入伪结的稳定性、动态性和展开 路径，以及结构。一系列的核磁共振实验将是 用来详细研究RNA的这些动态方面 假结。

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.