Global Analysis of RNA-protein Interactions in Plants

智力上的功绩。基因表达(或转录)的产物，RNA和这些RNA的产物(通过翻译)，即蛋白质，经常相互作用，从而允许许多转录后机制来控制mRNA的表达和翻译。事实上，从转录本在细胞核中的最初合成到最终在细胞质中的翻译和衰退，mRNAs参与了与数百个RNA结合蛋白(RBP)的大量复杂相互作用，导致动态多组分核糖核蛋白复合体(RNPs)的形成。研究表明，这些RNP对植物生物学的重要性已经被证明，植物的发育、激素和胁迫反应需要许多在植物转录组中具有不同功能和调控靶点的限制性商业惯例。事实上，过量或低表达特定限制性商业惯例的作物可以使其对某些生物和非生物胁迫更具抵抗力。因此，RNA-蛋白质相互作用对于植物细胞中许多RNA分子的功能、加工和调节是必要的。不幸的是，关于植物中RNA-蛋白质相互作用的全球格局仍然知之甚少。为此，该项目将使用一种新的技术来揭示这些RNA-蛋白质相互作用以及它们在激素和胁迫反应过程中对植物转录组和翻译组的影响。这项研究还将开始揭示植物王国中RNA-蛋白质相互作用位点的保守。总体而言，该项目将为RNA-蛋白质相互作用在调控真核基因表达中的作用提供前所未有的见解，并为未来专注于作物改良和以植物为重点的生物燃料研究提供重要的新发现和资源。这个项目将对生物学研究产生重大影响，对课堂也是如此。具体地说，这个项目的一个关键特点是它为研究生、本科生和高中生提供了许多研究机会。此外，这项研究获得的数据不仅将为其他科学家提供资源，而且还将用于RNA生物学课程，以开发侧重于核糖核蛋白复合体的教学模块，并用于基于计算的高通量测序数据分析，用于基因组科学课程。这些教学模块将免费提供，以便其他机构的教职员工和学生可以使用。这些免费提供的教学资源也将适用于高中水平的学习。总体而言，该项目将对未来科学家的培训和发展产生广泛影响，重点是计算机辅助研究方法。

Intellectual Merit. The product of gene expression (or transcription), RNA and the product of these RNAs (via translation), namely proteins, often interact to allow numerous post-transcriptional mechanisms to control mRNA expression and translation. In fact, from the initial synthesis of transcripts in the nucleus to their final translation and decay in the cytoplasm, mRNAs are involved in a multitude of complex interactions with hundreds of RNA binding proteins (RBPs), resulting in the formation of dynamic multi-component ribonucleoprotein complexes (RNPs). The importance of these RNPs to plant biology has been demonstrated by studies revealing that plant developmental, hormone, and stress responses require many RBPs with diverse functions and regulatory targets within the plant transcriptome. In fact, crop plants over- or under-expressing specific RBPs can be made more resistant to certain biotic and abiotic stresses. Thus, RNA-protein interactions are necessary for the functionality, processing, and regulation of many RNA molecules in plant cells. Unfortunately, there is still very little known about the global landscape of RNA-protein interactions in plants. To this end, this project will use a novel technology to uncover these RNA-protein interactions and their effect on the plant transcriptome and translatome during hormone and stress responses. This research will also begin to reveal the conservation of RNA-protein interaction sites within the plant kingdom. Overall, this project will provide unprecedented insights into the role of RNA-protein interactions in regulating eukaryotic gene expression, as well as provide important new findings and resources for future studies focused on crop improvement and plant-focused biofuel research.Broader Impacts. This project will have a significant impact on biological research, and the classroom as well. Specifically, a key feature of this project is that it provides many research opportunities for graduate, undergraduate, and high school students. Furthermore, the data obtained from this research will provide a resource not only to other scientists, but also for use in an RNA Biology course to develop teaching modules focused on ribonucleoprotein complexes and for computational-based analyses of high-throughput sequencing data for use in Genome Sciences courses. These teaching modules will be made freely available, so that faculty and students at other institutions can make use of them. These freely available teaching resources will also be made appropriate for high school level learning. Overall, this project will have a broad impact on the training and development of future scientists, with a focus on computer-aided research methods.