Structural Studies of U6 snRNA

描述（申请人提供）：准确有效的核前mRNA 剪接是真核生物的基本要素。异常的前体mRNA剪接已经被 与许多疾病有关。这些包括乳腺癌、结肠直肠癌、上皮癌 和卵巢癌，以及神经退行性疾病，如帕金森氏症 和老年痴呆症我们的目标是发展一个原子级的理解， 前体mRNA剪接的关键成分的结构和功能 U6 snRNA的结构。目前，我国严重缺乏结构性 关于U6 snRNA 我们假设U6 snRNA结构的功能是与镁离子配位， 剪接过程中离子辅因子，且不需要蛋白质和其它RNA 用于镁结合。我们将使用核磁共振光谱来研究 U6 snRNA的结构、功能、动力学和金属结合特性。我们 预计这项研究将为解释 过去对U6 snRNA该区域汇编的遗传和生化数据 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 为了验证我们的假设，即U6茎环足以协调 催化必需的金属离子。2.确定结构和 研究了S.酿酒酵母U6 snRNA分子内茎环 通过核磁共振。相应的人类U6和U6atac结构也将被 研究了3.研究U6突变表型的结构基础。我们 将把构造效应与研究得很好的S.酿酒酵母突变株 表型4.研究周围U6 snRNA序列的影响 剪接体蛋白Prp24位于U6 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.