Novel PT(II) Reagents for RNA Biochemistry

有了这个奖项，化学部门的生命过程化学项目将资助俄勒冈大学的Victoria DeRose博士开发新的金属基化合物，用于预测核糖核酸（RNA）生物聚合物的结构。后基因组时代的一个主要认识是细胞RNA通过不同机制控制基因表达的程度，其中许多机制仍在发现中。预测和确定大rna的结构是在追求阐明这些机制的重大挑战。小型化学探针是鉴定RNA结构元素的有力工具。本项目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。由于该项目需要综合使用合成、分子生物学以及分析和计算方法，因此它促进了化学不同分支之间的跨学科互动，并为本科生和研究生（包括女性）提供了一个劳动力培训环境，为STEM领域的职业生涯做准备。该计划的具体目标是开发方形平面，交换惰性Pt（II）配合物作为RNA交联试剂，并将其用作RNA结构表征的工具。Pt（II）化合物交联了富含嘌呤但非规范的RNA区域，如内部环和连接。Pt(II)-交联物种通过RNA分离方法是稳健的，但在温和条件下是可逆的。为了进行交联分析，将合成叠氮化物或炔修饰的Pt(II)-“Click”试剂，并安装手柄，用于Pt(II)- rna物种的选择性后处理修饰。将根据Pt（II）配合物的独特化学性质开发结构分析方案，包括高通量测序方法和高分辨率质谱分析。硫代膦“诱饵”配体将被纳入RNA中，为金属配合物创造新的交联位点。提高Pt（II）试剂用于RNA结构分析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多核Pt（II）化合物的测试。本文描述的基于Pt（II）的交联方法有可能成为RNA结构预测和细胞内小分子RNA可及性探针的工具。

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.