Interactions of Transition Metal Ions with RNA: Structure and Function

该奖项由化学部的生命过程化学项目资助。俄勒冈大学的Victoria J. DeRose教授研究了复杂核糖核酸（RNA）生物聚合物的结构。RNA参与各种各样的生命维持过程。它采用了复杂和动态的结构，用现有的方法来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作为RNA结构探针，并用于测定细胞中的RNA结构。本项目的成果是RNA结构分析的新方法和试剂，以及细胞中大RNA结构的新信息。该项目采用了广泛的方法，为劳动力发展提供了独特的跨学科培训环境，包括为女性和其他未被充分代表的科学、技术、工程和数学（STEM）群体提供夏季本科培训。本研究的目标是开发大RNA中远距离三级相互作用的方法，并了解靶向化合物到复杂RNA结构的决定因素。该项目开发交换惰性铂（Pt(II)）配合物作为RNA交联试剂，并将其纳入RNA结构测定方法中。Pt（II）化合物交联了RNA中富含嘌呤但非规范的区域，如内环和连接，并且通过RNA分离方法具有鲁棒性。为了进行交联分析，合成了新型修饰Pt（II）点击试剂，并安装了手柄，用于Pt(II) RNA物种的选择性后处理修饰。基于这些Pt（II）配合物的独特化学性质和高通量测序方法的结构分析方案，以及直接对交联物种进行成像的方法被开发出来。通过设计增强的Pt配体来探索小分子RNA相互作用的特异性。这些方法用于预测体外和细胞内rna的结构，包括长链非编码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.