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: 10210410
• 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/10210410
• 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.

项目摘要 近年来，人们对RNA三螺旋的兴奋迅速增长，这是由于它们在生物学中的新发现。 端粒合成，前体mRNA剪接，保护促癌RNA免受降解，现在是基因组 编辑.因此，发现和表征更多的RNA三螺旋是研究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。11自然 迄今为止，已经验证了存在的RNA三螺旋，并且预测会有更多，因此显然需要 开发分子工具，以确定整个自然界的RNA三螺旋的全球景观。 在此，主要研究者建议描述MALAT 1（转移相关肺转移）的特征。 腺癌转录物1）RNA三螺旋及其蛋白结合配偶体，胃L16（甲基转移酶样 蛋白质16）。MALAT 1三螺旋和胃L16复合物是第一个假定的三链 核糖核蛋白复合物，从而代表了一个独特的机会，提供前所未有的见解， 三链RNA生物学在研究领域#1和#2中，研究了胃L16和小 将首次确定特异性识别MALAT 1三螺旋并与之相互作用的分子 结合X射线晶体学和生物化学方法。这项工作将产生 开发新的实验工具以在全球发现更多RNA所需的基础知识 三重螺旋此外，发现了一种全新的蛋白质，即三链RNA结合蛋白， 蛋白质，预计。在研究领域#3和#4中，人胃L16作为推定的胃蛋白酶的生物学功能被证实是一种重要的生物学功能。 将使用体外和基于细胞的生物化学测定来研究三链RNA结合蛋白。 这项功能研究将集中在胃L16-MALAT 1三螺旋复合物的作用以及 胃L16及其RNA结合伴侣在Miller-Dieker综合征（一种罕见的 其中mettl 16基因的一个染色体拷贝缺失的神经变性疾病。因此，L16 和MALAT 1三重螺旋代表了一个模型系统，具有很高的潜力，以改变三重螺旋领域， 单链RNA生物学，开启了生物医学研究的未知领域。