Interactions of Transition Metal Ions with RNA: Structure and Function

化学部的生命过程化学项目为该奖项提供资金。来自俄勒冈州大学的维多利亚J. DeRose教授研究复杂核糖核酸（RNA）生物聚合物的结构。RNA参与各种各样的生命维持过程。它采用复杂和动态的结构，用当前的方法进行监测和预测具有挑战性。在这项研究中，开发了新的铂化合物作为RNA结构探针，并用于确定细胞中的RNA结构。该项目的成果是用于RNA结构分析的新方法和试剂，以及关于细胞中大RNA结构的新信息。该项目采用的方法广泛，为劳动力发展提供了独特的跨学科培训环境，包括为妇女和其他代表性不足的群体提供科学、技术、工程和数学（STEM）方面的夏季本科生培训。本研究的目标是开发大RNA中远程三级相互作用的方法，并了解将化合物靶向复杂RNA结构的决定因素。该项目开发了交换惰性铂（Pt（II））络合物作为RNA交联试剂，并将其纳入RNA结构测定方法中。Pt（II）化合物交联RNA中富含嘌呤但非典型的区域，例如内部环和接头，并且通过RNA分离方法是稳健的。为了能够进行交联分析，合成了新型修饰的Pt（II）点击试剂，其中安装了用于Pt（II）RNA种类的选择性处理后修饰的手柄。基于这些Pt（II）络合物的独特化学性质和高通量测序方法的结构分析方案被开发，沿着直接成像交联物质的方法。小分子RNA相互作用的特异性通过设计增强的Pt配体来探测。这些方法用于预测体外和细胞内RNA的结构，包括长非编码RNA的三级结构。

The Chemistry of Life Processes Program in the Chemistry Division funds this award. Professor Victoria J. DeRose from the University of Oregon investigates the structures of complex ribonucleic acid (RNA) biopolymers. RNA takes part in a wide variety of life sustaining processes. It adopts complex and dynamic structures that are challenging to monitor and predict with current methods. In this research, new platinum compounds are developed as RNA structure probes and used to determine RNA structures in cells. The outcomes of this project are novel methods and reagents for RNA structure analysis, and new information about large RNA structures in cells. The breadth of methods applied to this project supports a unique interdisciplinary training environment for workforce development, including summer undergraduate training for women and other underrepresented groups in science, technology, engineering and mathematics (STEM). The goals of this research are to develop methods for long-range tertiary interactions in large RNAs and to understand determinants for targeting compounds to complex RNA structures. The project develops exchange-inert platinum (Pt(II)) complexes as RNA crosslinking reagents and incorporates them into approaches for RNA structure determination. Pt(II) compounds crosslink purine-rich but noncanonical regions in RNA such as internal loops and junctions and are robust through RNA isolation methods. To enable crosslink analysis, novel modified Pt(II) click reagents are synthesized with installed handles for selective post-treatment modification of Pt(II) RNA species. Structure analysis protocols based on the unique chemistry of these Pt(II) complexes and high-throughput sequencing methods are developed, along with methods to directly image crosslinked species. The specificity of small-molecule RNA interactions is probed through design of enhanced Pt ligands. These approaches are used to predict structures of RNAs in vitro and in-cell, including tertiary structures in long non-coding RNAs.