Characterization of a Model System to Advance Triple-Stranded RNA Biology

推进三链 RNA 生物学的模型系统的表征

• 批准号: 9797666
• 负责人: JESSICA Ann BROWN
• 金额: $ 38.63万
• 依托单位: UNIVERSITY OF NOTRE DAME
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9797666
• 关键词: Area; Binding Proteins; Biochemical; Biological; Biological Assay; Biological Models; Biological Process; Biology; Cells; Complex; Development; Disease; Foundations; Genes; Health; Human; In Vitro; Investigation; Knowledge; MALAT1 gene; Malignant Neoplasms; Mediating; Methods; Methyltransferase; Miller-Dieker Syndrome; Molecular; Nature; Neurodegenerative Disorders; Principal Investigator; Proteins; RNA; RNA Binding; RNA Degradation; RNA Splicing; RNA-Binding Proteins; Research; Ribonucleoproteins; Role; Structure; X-Ray Crystallography; base; design; genome editing; insight; mRNA Precursor; novel; small molecule; telomere; time use; tool; triple helix

Project Summary Excitement about RNA triple helices has grown rapidly in recent years due to their newfound biological roles in telomere synthesis, pre-mRNA splicing, protecting cancer-promoting RNAs from degradation and now genome editing. Therefore, the discovery and characterization of more RNA triple helices is a pre-requisite to elucidating other fundamental biological processes mediated by triple-helical structures. As only 11 naturally occurring RNA triple helices have been validated to date and more are predicted, there is a clear need for the development of molecular tools to determine the global landscape of RNA triple helices throughout nature. Herein, the principal investigator proposes to characterize the MALAT1 (metastasis-associated lung adenocarcinoma transcript 1) RNA triple helix and its protein-binding partner, METTL16 (methyltransferase-like protein 16). The MALAT1 triple helix and METTL16 complex is the first putative triple-stranded ribonucleoprotein complex, thereby representing a unique opportunity to provide unprecedented insights into triple-stranded RNA biology. In Research Areas #1 and #2, the structural basis of how METTL16 and small molecules specifically recognize and interact with the MALAT1 triple helix will be determined for the first time using a combination of X-ray crystallography and biochemical methods. This effort will generate the foundational knowledge needed to develop novel experimental tools for the global discovery of more RNA triple helices. Moreover, the discovery of an entirely new class of proteins, that is triple-stranded RNA-binding proteins, is anticipated. In Research Areas #3 and #4, the biological function of human METTL16 as a putative triple-stranded RNA-binding protein will be investigated using both in vitro and cell-based biochemical assays. This functional investigation will focus on the roles of the METTL16-MALAT1 triple helix complex as well as broader roles for METTL16 and its RNA-binding partners in the context of Miller-Dieker Syndrome, a rare neurodegenerative disease in which one chromosomal copy of the mettl16 gene is deleted. Thus, METTL16 and the MALAT1 triple helix represent a model system with high potential to transform the field of triple- stranded RNA biology, opening unforeseen areas of biomedical investigation.

项目概要 近年来，人们对 RNA 三螺旋的兴趣迅速增长，因为它们在 端粒合成、前 mRNA 剪接、保护促癌 RNA 免遭降解，现在是基因组 编辑。因此，更多RNA三螺旋的发现和表征是研究RNA三螺旋的先决条件。 阐明由三螺旋结构介导的其他基本生物过程。自然只有11个 迄今为止，已经验证了发生的 RNA 三螺旋，并且预测了更多，因此显然需要 开发分子工具来确定整个自然界中 RNA 三螺旋的全球景观。 在此，主要研究者建议表征 MALAT1（转移相关肺） 腺癌转录本 1) RNA 三螺旋及其蛋白结合伴侣 METTL16（甲基转移酶样 蛋白质 16)。 MALAT1 三螺旋和 METTL16 复合物是第一个假定的三链 核糖核蛋白复合物，从而提供了一个独特的机会来提供前所未有的见解 三链RNA生物学。在研究领域#1和#2中，METTL16和小型的结构基础 特异性识别MALAT1三螺旋并与之相互作用的分子将首次被确定 结合使用 X 射线晶体学和生化方法。这项努力将产生 开发新的实验工具以在全球范围内发现更多 RNA 所需的基础知识 三重螺旋。此外，发现了一类全新的蛋白质，即三链 RNA 结合蛋白 蛋白质，是预料之中的。在研究领域 #3 和 #4 中，人类 METTL16 的生物学功能作为假定的 将使用体外和基于细胞的生化测定来研究三链RNA结合蛋白。 这项功能研究将重点关注 METTL16-MALAT1 三螺旋复合物的作用以及 METTL16 及其 RNA 结合伴侣在米勒-迪克综合征（一种罕见的罕见疾病）的背景下发挥更广泛的作用 一种神经退行性疾病，其中 metl16 基因的一个染色体副本被删除。因此，METTL16 MALAT1 三螺旋代表了一个具有很大潜力改变三螺旋领域的模型系统 搁浅RNA生物学，开启了生物医学研究不可预见的领域。