RESEARCH-PGR: Defining the Sumoylation System in Maize and Its Roles in Stress Protection

研究-PGR：定义玉米中的苏酰化系统及其在应激保护中的作用

• 批准号: 1546862
• 负责人: Richard Vierstra
• 金额: $ 93.01万
• 依托单位: Washington University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-15 至 2020-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1546862&HistoricalAwards=false
• 关键词: RESEARCH; PGR; Defining; Sumoylation; System

Crop plants require suitable mechanisms to survive and perform well under unfavorable environments in order to maintain high productivity. Recently, a small protein called SUMO was discovered, and it appears to protect plants against environmental stresses like heat and drought. Recent experiments with plants exposed to environmental stresses have shown that the SUMO protein may significantly affect how genes are expressed under those unfavorable conditions. The SUMO protein becomes rapidly and reversibly attached to a group of proteins that affect the pattern of gene expression during environmental stress. The goals of this project are to understand the functions of SUMO during stress and determine whether its manipulation in crops might provide novel approaches to improve stress tolerance. This project will characterize the mode of action of the SUMO protein, and identify all the other proteins that interact with SUMO under stress. To help understand how SUMO helps maize survive stress, we will also determine why maize mutants missing key components are now hypersensitive to stress. Collectively, this fundamental research will identify key points in maize SUMOylation that can be exploited to improve stress protection, not only in maize but in other crops as well. This project will train postdocs, graduate students, and undergraduates at Washington University in St. Louis. It will also develop patentable technologies related SUMO and stress tolerance that can sustainably enhance food and biofuel crop yield.SUMO is an influential regulator in eukaryotes that works following its post-translation addition to other intracellular proteins. Recent studies with Arabidopsis discovered that SUMOylation selectively modifies a number of critical regulators during stress that impact chromatin accessibility, DNA/histone modification, transcription, nuclear pore function, and mRNA processing/export. Their combined activities imply that stress-induced SUMOylation helps plants reversibly adjust chromatin architecture and the resulting RNA landscape to better survive adverse conditions. Unfortunately, appreciation of SUMO is lacking for crops, thus precluding rational redesign for agricultural benefit. To overcome this knowledge gap, this project will define how SUMOylation works in maize (Zea mays), using as essential backdrops our recent success in creating germplasm to study SUMO via proteomic approaches. This project will: (i) describe how the maize SUMO system works biochemically and responds to various environmental challenges, (ii) combine transgenic lines expressing tagged SUMOs with established mass spectrometric methods to define the maize 'SUMOylome' and its linkage sites, (iii) determine quantitatively how the SUMOylation status of individual targets is impacted by stress, and (iv) understand how SUMO helps maize survive adverse environments through the phenotypic and biochemical analyses of system mutants generated by either Mu transposition or CRISPR/Cas9 gene editing technologies. Collectively, this project will generate much-needed reagents, techniques, and mutants that will help define how SUMO reorganizes maize chromatin and its transcriptome during stress, and identify key points in SUMOylation that can be manipulated to improve stress protection in many crop species.

作物需要合适的机制来在不利的环境下生存和表现良好，以保持高生产力。最近，一种名为SUMO的小蛋白被发现，它似乎可以保护植物免受高温和干旱等环境胁迫。最近对暴露于环境胁迫的植物进行的实验表明，SUMO蛋白可能会显着影响基因在这些不利条件下的表达。SUMO蛋白快速可逆地附着在一组影响环境胁迫期间基因表达模式的蛋白质上。该项目的目标是了解SUMO在胁迫期间的功能，并确定其在作物中的操纵是否可能提供新的方法来提高胁迫耐受性。 该项目将描述SUMO蛋白的作用模式，并确定在压力下与SUMO相互作用的所有其他蛋白质。为了帮助理解SUMO如何帮助玉米在压力下生存，我们还将确定为什么缺失关键成分的玉米突变体现在对压力高度敏感。 总的来说，这项基础研究将确定玉米SUMO化的关键点，这些关键点不仅可以用于玉米，还可以用于其他作物。该项目将在圣路易斯的华盛顿大学培养博士后、研究生和本科生。它还将开发与SUMO和抗逆相关的专利技术，可持续提高粮食和生物燃料作物的产量。SUMO是真核生物中一种有影响力的调节剂，在翻译后添加到其他细胞内蛋白质中。 最近对拟南芥的研究发现，SUMO化在胁迫期间选择性地修饰许多关键调节子，这些调节子影响染色质可及性、DNA/组蛋白修饰、转录、核孔功能和mRNA加工/输出。 它们的组合活性意味着胁迫诱导的SUMO化有助于植物可逆地调整染色质结构和由此产生的RNA景观，以更好地在不利条件下生存。 不幸的是，相扑的赞赏是缺乏对作物，从而排除了合理的重新设计农业利益。 为了克服这一知识差距，该项目将定义SUMO化如何在玉米（玉米）中起作用，使用我们最近成功创建种质以通过蛋白质组学方法研究SUMO的重要背景。 该项目将：（i）描述玉米SUMO系统如何在生物化学上起作用并响应各种环境挑战，（ii）将表达标记的SUMO的联合收割机转基因系与已建立的质谱方法组合以定义玉米“SUMO 1 ome”及其连锁位点，（iii）定量确定单个靶标的SUMO化状态如何受到胁迫的影响，以及（iv）了解SUMO如何通过对Mu转座或CRISPR/Cas9基因编辑技术产生的系统突变体进行表型和生化分析，帮助玉米在不利环境中生存。 总的来说，该项目将产生急需的试剂，技术和突变体，这将有助于确定SUMO如何在胁迫期间重组玉米染色质及其转录组，并确定SUMO化中的关键点，这些关键点可以被操纵以改善许多作物物种的胁迫保护。