Structural Studies of U6 snRNA

描述(申请人提供)：准确有效的核前基因 剪接对于真核生物来说是必不可少的。异常的前信使核糖核酸剪接 与许多疾病有关。包括乳房、结直肠、上皮组织 和卵巢癌，以及帕金森氏症等神经退行性疾病 和阿尔茨海默氏症。我们的目标是在原子水平上理解 前信使核糖核酸剪接的一个关键成分的结构和功能 机械，U6小核糖核酸。目前，严重缺乏结构性的 有关U6单链RNA的信息。 我们假设U6SnRNA结构具有与镁离子配位的功能 剪接过程中离子辅助因子，不需要蛋白质和其他RNA 用于镁的结合。我们将使用核磁共振波谱来研究 U6单链RNA的结构、功能、动力学和金属结合特性。我们 预期这项研究将为解释 过去收集的关于U6单链RNA这个区域的遗传和生化数据 十年。我们的具体目标是： 1.定义金属离子与U6 SnRNA茎环的相互作用。我们将使用核磁共振 为了测试我们的假设，即U6茎环足以协调 催化必需的金属离子。2.确定结构和 酿酒酵母U6单链RNA分子内茎环的动力学研究 核磁共振仪。相应的人类U6和U6atac结构也将是 调查过了。3.研究U6突变表型的结构基础。我们 将结构效应与研究充分的酿酒酵母突变株相关联 表型。4.研究周围的U6单链RNA序列和 剪接体蛋白Prp24位于U6的SnRNA半环结构上。

DESCRIPTION (provided by applicant): Accurate and efficient nuclear pre-mRNA splicing is essential to eukaryotic life. Aberrant pre-mRNA splicing has been linked to a number of diseases. These include breast, colorectal, epithelial and ovarian cancer, as well as neurodegenerative diseases such as Parkinson's and Alzheimer's. Our objective is to develop an atomic-level understanding of the structure and function of a key component of the pre-mRNA splicing machinery, that of U6 snRNA. Currently, there is a profound lack of structural information about U6 snRNA. We hypothesize that the U6 snRNA structure functions to coordinate a magnesium ion cofactor during splicing, and that proteins and other RNAs are not required for magnesium binding. We will use NMR spectroscopy to investigate the structure, function, dynamics and metal binding properties of U6 snRNA. We anticipate that this research will provide a basis for the interpretation of genetic and biochemical data compiled on this region of U6 snRNA over the past decade. Our specific aims are: 1. Define metal ion interactions with the U6 snRNA stem-loop. We will use NMR to test our hypothesis that the U6 stem-loop is sufficient for coordination of the catalytically essential metal ion. 2. Determine the structure and investigate the dynamics of the S. cerevisiae U6 snRNA intramolecular stem-loop by NMR. The corresponding human U6 and U6atac structures will also be investigated. 3. Investigate the structural basis for U6 mutant phenotypes. We will correlate structural effects with well studied S. cerevisiae mutant phenotypes. 4. Investigate the influence of surrounding U6 snRNA sequences and the splicoesomal protein Prp24 on the U6 snRNA sem-loop structure.