Novel PT(II) Reagents for RNA Biochemistry

通过这一奖项，化学部生命过程化学计划资助俄勒冈大学的Victoria DeRose博士开发用于预测核糖核酸(RNA)生物聚合物结构的新型金属化合物。后基因组时代的一个主要认识是细胞RNA通过不同的机制控制基因表达的程度，其中许多机制仍在被发现。预测和确定大RNA的结构是阐明这些机制的重大挑战。小的化学探针可以成为识别RNA结构元素的强大工具。本项目中开发的铂基化合物和方法旨在为高通量RNA结构预测提供这样的新工具。由于这个项目需要使用合成、分子生物学以及分析和计算方法的组合，它促进了不同化学分支之间的跨学科互动，并为准备在STEM领域就业的本科生和研究生，包括女性，提供了一个劳动力培训环境。该计划的具体目标是开发方形平面、交换惰性铂(II)络合物作为RNA交联剂，并将其用作RNA结构表征的工具。铂(II)化合物交联型富含嘌呤但非正则的RNA区域，如内环和连接。铂(II)-交联物种通过RNA分离方法是健壮的，但在温和的条件下是可逆的。为了能够进行交联分析，将合成叠氮或炔修饰的铂(II)-“Click”试剂，并安装手柄以选择性地对铂(II)-RNA物种进行后处理修饰。将根据铂(II)络合物的独特化学特性开发结构分析方案，包括高通量测序方法和高分辨率质谱学。硫代硫酸盐“诱饵”配体将被结合到RNA中，为金属络合物创造新的交联点。加强用于RNA结构分析的铂(II)试剂的性能的努力将包括用更长的连接体修饰可点击的单核化合物和测试多核铂(II)化合物。这里描述的基于铂(II)的交联法有可能成为RNA结构预测的工具和小分子可及性的细胞内探针。

With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Victoria DeRose from the University of Oregon to develop new metal-based compounds for predicting the structure of ribonucleic acid (RNA) biopolymers. A major realization in the post-genomic era is the extent to which cellular RNA governs gene expression through different mechanisms, many of which are still being discovered. The prediction and determination of the structure of large RNAs are significant challenges in the pursuit of the elucidation of these mechanisms. Small chemical probes can be powerful tools for the identifying structural elements of RNA. The platinum-based compounds and methods developed in this project are designed to provide such new tools for high-throughput RNA structure prediction. As this project requires use of a combination of synthesis, molecular biology, and analytical and computational methods, it fosters interdisciplinary interactions between different branches of chemistry, and it provides a workforce training environment for undergraduate and graduate students, including women, who prepare for careers in STEM fields. The specific goals of this program are to develop square-planar, exchange-inert Pt(II) complexes as RNA crosslinking reagents and use them as tools for RNA structure characterization. Pt(II) compounds crosslink purine-rich but noncanonical regions of RNA, such as internal loops and junctions. Pt(II)-crosslinked species are robust through RNA isolation methods but reversible in mild conditions. To enable crosslink analysis, azide- or alkyne-modified Pt(II)-"Click" reagents with installed handles for selective post-treatment modification of Pt(II)-RNA species will be synthesized. Structure analysis protocols will be developed based on the unique chemistry of the Pt(II) complexes, including high-throughput sequencing methods and high-resolution mass spectrometry. Phosphorothioate "bait" ligands will be incorporated in RNA to create new crosslinking sites for the metal complexes. Efforts to enhance the properties of Pt(II) reagents for RNA structure analysis will include modification of clickable mononuclear compounds with longer linkers and testing of multinuclear Pt(II) compounds. The Pt(II)-based crosslinking methods described here have the potential to become tools for RNA structure prediction and in-cell probes of RNA accessibility to small molecules.