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.

作物工厂需要适当的机制才能生存并在不利的环境下表现良好，以维持高生产率。最近，发现了一种称为Sumo的小蛋白质，它似乎可以保护植物免受热和干旱等环境压力。对暴露于环境应力的植物的最新实验表明，相扑蛋白可能会显着影响基因在那些不利条件下的表达方式。相扑蛋白在影响环境应激过程中影响基因表达模式的一组蛋白质上迅速和可逆地附着。该项目的目标是了解压力期间Sumo的功能，并确定其在农作物中的操作是否可以提供提高压力耐受性的新方法。 该项目将表征相扑蛋白的作用方式，并确定在压力下与Sumo相互作用的所有其他蛋白质。为了帮助了解相扑如何帮助玉米生存压力，我们还将确定为什么缺少关键成分的玉米突变体现在对压力过敏。 总的来说，这项基本研究将确定可以利用玉米sumoylation的关键点，不仅在玉米中，而且在其他农作物中也可以改善压力保护。该项目将在圣路易斯华盛顿大学培训博士后，研究生和本科生。它还将开发可持续的相关技术和应力耐受性，从而可持续增强食物和生物燃料作物的产量。Sumo是真核生物中有影响力的调节剂，其在升级后加入其他细胞内蛋白质后起作用。 最近对拟南芥的研究发现，在压力期间，Sumoylation选择性地修饰了许多关键调节剂，从而影响染色质的可及性，DNA/组蛋白修饰，转录，核孔功能和mRNA处理/导出。 他们的合并活动表明，压力诱导的sumoylation有助于植物可逆地调节染色质结构和所得的RNA景观，从而更好地生存不良条件。 不幸的是，缺乏对农作物的欣赏，从而排除了农业利益的理性重新设计。 为了克服这一知识差距，该项目将定义Sumoylation在玉米（Zea Mays）中的工作原理，并以我们最近在创造种质中通过蛋白质组学方法来研究Sumo的基本背景。 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通过MU转位或CRISPR/CAS9基因编辑技术产生的系统突变体的表型和生化分析。 总的来说，该项目将产生急需的试剂，技术和突变体，这些试剂，技术和突变体将有助于定义Sumo在压力期间如何重组玉米染色质及其转录组，并确定可以操纵的Sumoylation中的关键点，以改善许多作物的应激保护。