Analysis of the Ribonucleases of Arabidopsis thaliana

基因表达的变化对植物的生长、发育和抗病性至关重要。 更好地了解基因表达是如何调节的是植物工程过程的基础，因此，提高农业生产。 植物在多个水平上调节基因表达，本项目的目的是表征称为内切核糖核酸酶的蛋白质，其在mRNA降解水平上调节基因表达。 该项目的一个重要方面是确定这些内切核糖核酸酶如何影响植物的生物学，包括植物生长和发育。 该项目将对年轻科学家进行基因调控基本方面的培训。 这位年轻的科学家将通过与系统生物学和化学工程专家的合作者进行互动来接受跨学科的培训。 一个影子科学家计划将使中学生有机会会见科学家，参观研究实验室，并看到这个项目的实验正在进行中。 其他外联活动旨在吸引公众参与，激发来自各种背景和年龄组的人们对科学的兴趣。 该项目的长期目标是阐明参与拟南芥mRNA降解的核糖核酸酶（RNase）的作用。 研究核糖核酸内切酶在基因转录后调控中的作用是一个不发达的研究领域，但却是一个及时的领域。 这项研究提出的假设是，阐明核糖核酸内切酶的靶点和转录后影响将增强我们对植物mRNA衰变中关键参与者的理解，例如外泌体，以及鲜为人知或推定的核糖核酸内切酶，其作用尚未发现。 为了测试这个想法和其他想法，该项目将首先创建一组双突变体来稳定由内切核糖核酸酶产生的mRNA衰变中间体，以便它们可以被深度测序;将单个内切核糖核酸酶突变引入xrn 4无效突变体背景中以实现这一目标。 其次，为了鉴定内切核糖核酸酶的底物，将使用RNA末端和RNA-Seq数据的平行分析在双突变体和对照中在全球范围内分析RNA衰变中间体和RNA水平。 如果任何内切核糖核酸酶作用于无义介导的mRNA衰变（NMD）靶点，考虑到植物缺乏后生动物NMD内切核糖核酸酶SMG 6，这将是有趣的。 第三，新的生物学影响的内切核糖核酸酶将假设和测试的基础上，其切割基板的功能协会的单一突变体。 第四，将研究涉及核糖核酸内切酶的转录后机制。核糖核酸内切酶的定位，与它们相互作用的蛋白质的鉴定以及其他转录后数据将完善表型假设和模型。 除了利用虚拟植物来实现这一目标外，该项目还将通过新的转录后组件来增强其力量。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Changes in gene expression are crucial for plant growth, development and resistance to pathogens. A better understanding of how gene expression is regulated is fundamental to the process of plant engineering and hence to improved agricultural production. Plants regulate gene expression at multiple levels and the aim of this project is to characterize proteins called endoribonucleases, which regulate gene expression at the level of mRNA degradation. An important aspect of this project is to determine how these endoribonucleases impact the biology of plants, including plant growth and development. This project will train young scientists on fundamental aspects of gene regulation. The young scientist will receive interdisciplinary training through interactions with collaborators who are experts in systems biology and chemical engineering. A Shadow a Scientist program will give middle school students the chance to meet scientists, tour a research laboratory, and to see experiments for this project in progress. Other outreach activities aim to engage the public and to inspire scientific interest in people from a broad range of backgrounds and age groups. The long-term goal of this project is to elucidate the roles of ribonucleases (RNases) that participate in mRNA degradation in Arabidopsis. Examining the roles of endoribonucleases in post-transcriptional control of genes is an underdeveloped area of research, but a timely one. The hypothesis underlying this proposed research is that elucidating the targets and post-transcriptional impact of endoribonucleases will enhance our understanding of key players in plant mRNA decay, such as the exosome, as well as lesser known or putative endoribonucleases whose role is yet to be discovered. To test this idea and others, the project will first create a set of double mutants to stabilize mRNA decay intermediates produced by endoribonucleases so they can be deeply sequenced; single endoribonuclease mutations will be introduced into an xrn4 null mutant background to achieve this. Second, to identify substrates of the endoribonucleases, RNA decay intermediates and RNA levels will be analyzed on a global scale in the double mutants and controls using Parallel Analysis of RNA Ends and RNA-Seq data. Should any of the endoribonucleases act on nonsense-mediated mRNA decay (NMD) targets, that would be intriguing given that plants lack the metazoan NMD endoribonuclease, SMG6. Third, new biological impacts of the endoribonucleases will be hypothesized and tested in the single mutants on the basis of the functional associations of their cleaved substrates. Fourth, post-transcriptional mechanisms that involve the endoribonucleases will be investigated. Localization of the endoribonucleases, the identification of proteins interacting with them and other post-transcriptional data will refine phenotypic hypotheses and models. Beyond making use of The Virtual Plant for this pursuit, this project will enhance its power with new post-transcriptional components.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.