EAGER: Defining the SUMOylation System in Maize and its Roles in Stress Protection

EAGER：定义玉米中的 SUMOylation 系统及其在应激保护中的作用

• 批准号: 1232752
• 负责人: Richard Vierstra
• 金额: $ 30万
• 依托单位: University of Wisconsin-Madison
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-15 至 2016-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1232752&HistoricalAwards=false
• 关键词: EAGER; Defining; SUMOylation; System; Maize

The ability to detect and respond to stress is central to a plant's survival in a host of unfavorable environments and a key determinant of agricultural productivity under sub-optimal field conditions. Although a number of pathways have been described that confer specific protection to various abiotic and biotic challenges, a recent discovery of a potentially universal protective mechanism involving the Small Ubiquitin-related MOdifier (SUMO) may transform the current appreciation of stress biology. Specifically, it has been shown that the Arabidopsis SUMO polypeptide becomes covalently attached to numerous nuclear proteins and that the levels of these conjugates rise rapidly and reversibly after exposing plants to various abiotic stresses. Using novel quantitative proteomic approaches, it has been discovered that many of the SUMOylation targets are known critical regulators with their collective functions implying that SUMO addition engages a protective response that broadly alters chromatin accessibility, transcription, and mRNA processing/export. Taken together, these results suggest that SUMO might offer unique opportunities to globally manipulate the stress response for agricultural benefit. Unfortunately, the organization and functions of the SUMO system are largely unknown in other plant species, including all important agricultural crops, thus precluding rational redesign to improve crop plant productivity. Moreover, preliminary genome analyses of maize and rice revealed that the organization of the SUMO system in cereals might differ significantly from that in observed in Arabidopsis. This EAGER project proposes to define how SUMOylation works during stress in crops using maize (Zea mays) as the model. The specific aims are to: (i) delineate the SUMOylation system in maize using bioinformatic and biochemical methods and define kinetically how the system responds to stress; (ii) generate a library of maize mutants and transgenic lines affecting key components required for SUMO addition and release; (iii) define the "SUMOylome" of maize, quantify how the SUMOylation status of individual targets changes during stress and after recovery; and (iv) analyze SUMO pathway mutants phenotypically to determine how stress-induced SUMOylation may help maize survive adverse environments. Collectively, this project will generate much-needed tools and germplasm that can be exploited to understand how SUMO might reorganize maize chromatin and its transcriptome during stress, and identify key points in plant stress responses involving SUMOylation that can be manipulated for improved yield.The current understanding of SUMOylation in plants is still rudimentary and almost nonexistent in crop species where its manipulation may have substantial agricultural impact. This project will provide interdisciplinary training of the next generation of plant scientists working on crops. This research will collectively incorporate postdocs, graduate students, and undergraduates as well as high school students sponsored by the Wisconsin Youth Apprenticeship Program (YAP) in Biotechnology. During the course of this project, reagents, techniques, mutants, and transgenic lines will be generated that will provide a much needed foundation to investigate SUMOylation in maize, and hopefully offer new strategies to rationally alter the SUMO system for agricultural and medicinal benefit. Plant resources will be available through the Maize Genetics Cooperative Stock Center (http://maizecoop.cropsci.uiuc.edu). Raw and processed experimental data will be deposited into NCBI's Gene Expression Omnibus (GEO) and at Maize GDB (http://www.maizegdb.org/).

检测和应对压力的能力对于植物在不利的环境中的生存以及在亚最佳田间条件下的农业生产力的关键决定因素是至关重要的。尽管已经描述了许多途径，这些途径允许对各种非生物和生物挑战提供特定的保护，但最近发现涉及小型泛素相关修饰剂（SUMO）的潜在普遍保护机制（SUMO）可能会改变当前对应激生物学的欣赏。具体而言，已经表明，拟南芥Sumo多肽与众多核蛋白共价附着，并且这些结合物的水平在将植物暴露于各种非生物胁迫下后迅速而可逆地上升。使用新颖的定量蛋白质组学方法，已经发现许多Sumoylation靶标是已知的关键调节剂，其集体功能表明，Sumo添加的保护性响应可以广泛地改变染色质的可及性，转录和mRNA处理/导出。 综上所述，这些结果表明，相扑可能会提供独特的机会，以在全球操纵农业利益的压力反应。不幸的是，在其他植物物种（包括所有重要的农作物）中，相扑系统的组织和功能在很大程度上是未知的，因此排除了理性的重新设计以提高作物植物的生产率。此外，对玉米和水稻的初步基因组分析表明，谷物中的相扑系统的组织可能与拟南芥中观察到的相差明显不同。这个渴望的项目建议使用玉米（Zea Mays）作为模型来定义Sumoylation在农作物中的压力期间的工作方式。具体的目的是：（i）使用生物学和生化方法描绘玉米中的sumoylation系统，并在动力学上定义系统对压力的反应； （ii）生成一个玉米突变体和转基因线的库，影响相扑和释放所需的关键成分； （iii）定义玉米的“ sumoylome”，量化各个靶标在压力和恢复后的sumoylation状态如何变化； （iv）在表型上分析SUMO途径突变体，以确定应激诱导的Sumoylation如何帮助玉米生存不利的环境。总的来说，该项目将生成急需的工具和种质，可以利用这些工具和种质，以了解Sumo如何在压力期间重新组织玉米染色质及其转录组，并确定涉及Sumoylation的植物压力反应中的关键点，这些植物压力反应可以被操纵，这些植物应被操纵以提高植物的收益率。当前的植物的理解仍然是无与伦比的。该项目将提供跨学科的培训，对从事农作物的下一代植物科学家。这项研究将集体纳入博士后，研究生和本科生以及威斯康星州青年学徒计划（YAP）在生物技术中赞助的高中生。 在该项目过程中，将产生试剂，技术，突变体和转基因线，这将为调查玉米中的Sumoylation提供急需的基础，并希望提供新的策略，以合理地改变Sumo农业和药物益处的Sumo系统。 植物资源将通过玉米遗传学合作股票中心（http://maizecoop.cropsci.uiuc.edu）获得。 原始和加工的实验数据将沉积在NCBI的基因表达综合（GEO）和玉米GDB（http://www.maizegdb.org/）中。