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。来自俄勒冈大学的Victoria J.DeRose教授研究复杂的核糖核酸(RNA)生物聚合物的结构。RNA参与了多种维持生命的过程。它采用了复杂和动态的结构，用目前的方法进行监测和预测是具有挑战性的。在这项研究中，新的铂化合物被开发为RNA结构探针，并用于确定细胞中的RNA结构。该项目的成果是RNA结构分析的新方法和新试剂，以及关于细胞中大RNA结构的新信息。应用于该项目的各种方法为劳动力发展提供了独特的跨学科培训环境，包括为妇女和其他在科学、技术、工程和数学(STEM)方面代表性不足的群体提供暑期本科培训。这项研究的目标是开发在大RNA中进行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。该项目开发交换惰性铂(铂(II))络合物作为RNA交联剂，并将其纳入RNA结构确定的方法中。铂(II)化合物在RNA中的交联型富嘌呤但非正则区域，如内部环和连接，并且通过RNA分离方法是健壮的。为了能够进行交联分析，合成了新型的修饰的铂(II)点击试剂，并安装了手柄，用于对铂(II)RNA物种的选择性后处理修饰。基于这些铂(II)络合物的独特化学结构分析协议和高通量测序方法，以及直接成像交联物种的方法被开发出来。通过设计增强的铂配体，探讨了小分子RNA相互作用的特异性。这些方法被用来预测RNA的体外和细胞内结构，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.