NSF/MCB-BSF: RNA quality control in the chloroplast

优化植物性能的主要途径之一是提高它们收集太阳能量的能力。植物细胞含有叶绿体，光能在叶绿体中被收集并转化为糖，最终转化为食物、饲料和纤维。叶绿体包含自己的DNA和基因，光合作用必须正确地发挥作用。像所有的生物过程一样，这些功能容易出错，必须纠正这些错误才能使植物发挥作用。该项目旨在了解实现质量控制的基本而神秘的过程。这种类型的质量控制既不局限于叶绿体，也不局限于植物，因此本项目取得的进展将适用于其他生命系统。该项目还为培养实验和计算技术方面的本科生创造了机会，包括植物基因工程和生物信息学分析。学生们还将学习沟通技巧，并将接触到各种各样的职业选择。这种培训将通过指导和团队合作进行，这对早期职业科学家来说是重要的可转移技能。在叶绿体中，RNA质量控制是至关重要的，因为转录本身是相对不受调节的。该项目建立在拟南芥的生化、遗传和计算发现的基础上，重点研究基本的RNA质量控制酶RNase J， RNase J已被证明可以修剪5 ‘和3 ’ RNA末端，并消除有害的反义RNA。本项目旨在发现RNase J是如何被调控的，以及它的特异性是如何被赋予的。目前有两种主要的方法，第一种是建立转基因拟南芥系，表达具有信息性位点导向突变或缺失的RNaseJ版本，第二种是在体外生化测量RNaseJ的活性。总的来说，这些实验应该促进对叶绿体转录组如何形成的认识，并为在大量RNA分子中区分转录本的机制提供见解。该奖项是由分子和细胞生物科学部的遗传机制项目和生物科学理事会综合有机体系统部的植物基因组研究项目共同资助的。这个美国/以色列合作项目由美国国家科学基金会和以色列两国科学基金会支持。该奖项反映了美国国家科学基金会的法定使命，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。

One of the main avenues to optimizing plant performance is to improve their ability to harvest the sun’s energy. Plant cells contain chloroplasts where light energy is harvested and converted into sugars and ultimately into food, feed and fiber. The chloroplast contains its own DNA and genes that must function correctly for photosynthesis to occur. Like all biological processes, these functions are prone to errors that must be corrected for plants to function. This project seeks to understand the fundamental yet mysterious process by which this quality control is accomplished. Quality control of this type is not specific to chloroplasts, nor to plants, and therefore advances made in this project will be applicable to other living systems. The project also creates opportunities for training diverse undergraduates in experimental and computational techniques, including plant genetic engineering and bioinformatic analysis. Students will also be taught communications skills and will be exposed to a variety of career options. This training will occur through mentoring and teamwork, which are important transferable skills for early career scientists. In chloroplasts, RNA quality control is of paramount importance because transcription itself is relatively unregulated. This project builds on biochemical, genetic and computational findings in Arabidopsis, focusing on the essential RNA quality control enzyme, RNase J, which has been shown to trim 5’ and 3’ RNA termini and to eliminate deleterious antisense RNA. This project seeks to discover how RNase J is regulated, and how its specificity is imparted. The two major approaches are first, to create transgenic Arabidopsis lines that express versions of RNase J with informative site-directed mutations or deletions, and second, to biochemically measure the activities of RNaseJ in vitro. Overall, the experiments should advance knowledge of how the chloroplast transcriptome is shaped, and provide insights into mechanisms that discriminate transcripts among large populations of RNA molecules.This award was co-funded by the Genetic Mechanisms program of the Division of Molecular and Cellular Biosciences, and the Plant Genome Research Program of the Division of Integrative Organismal Systems in the Biological Sciences Directorate.This collaborative US/Israel project is supported by the US National Science Foundation and the Israeli Binational Science Foundation.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.