权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

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

基本信息

批准号：
10669618
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
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-01 至 2025-07-31
项目状态：
未结题

项目摘要

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，这是因为它们在端粒合成，前mRNA剪接，保护致癌RNA不被降解，现在是基因组正在编辑。因此，更多的RNA三螺旋的发现和鉴定是实现阐明由三螺旋结构介导的其他基本生物学过程。因为自然只有11个人到目前为止，出现的RNA三螺旋已经得到验证，并预测会有更多，显然需要开发分子工具以确定整个自然界的RNA三螺旋的全球格局。在此，首席研究人员建议将MALAT1(转移相关肺腺癌转录本1)RNA三螺旋及其蛋白结合伙伴--甲基转移酶样蛋白蛋白质16)。MALAT1三螺旋和METTL16复合体是第一个推测的三链核糖核蛋白复合体，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Small的结构基础首次确定了与MALAT1三螺旋特异识别和相互作用的分子使用X射线结晶学和生化方法相结合的方法。这一努力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，发现了一类全新的蛋白质，即三链RNA结合蛋白质，是预期的。在研究领域#3和#4中，人类METTL16作为一种推定的生物功能将使用体外和基于细胞的生化分析来研究三链RNA结合蛋白。这项功能研究将集中在METTL16-MALAT1三螺旋复合体的作用以及在罕见的Miller-Dieker综合征的背景下，METTL16及其RNA结合伙伴的更广泛的作用一种神经退行性疾病，指的是mettl16基因的一个染色体拷贝缺失。因此，METTL16 而MALAT1三螺旋代表了一个具有很高潜力的模型系统，可以改变三重螺旋的领域。搁浅的RNA生物学，打开了生物医学研究的未知领域。