SGER: Electrophile Responsive Proteome and Their Role in Plant Signaling Pathways

在动物系统中，各种环境和发育刺激会产生反应性亲电物质（RES），它们通过共价加合物的形成来修饰蛋白质，并引发适应性细胞反应。虽然这种被称为亲电介质介导的硫醇结合（EMTB）的翻译后修饰已知可调节动物的信号转导途径，但在植物系统中尚未报道。近年来研究发现，植物氧化脂素通路中一个关键酶的EMTB修饰是植物逆境应答的重要信号。该项目的目标是研究EMTB修饰在控制植物信号通路中的一般作用。智力优势：跨学科的方法将用于在全基因组范围内鉴定被胁迫诱导的12-氧植物二烯酸（12-OPDA）共价修饰的细胞靶蛋白，12-OPDA是植物中的一种脂质RES。12-OPDA结合蛋白靶点的一个子集将被充分表征并检查它们在植物胁迫信号通路中的作用。最后，将确定12-OPDA与其他应激诱导的硫反应性亲电试剂在蛋白质共价修饰方面的特异性。更广泛的影响：这项研究的结果将为一个新的、以前未被认识到的翻译后调控机制提供重要的基础见解，该机制控制着植物中一个复杂而关键的信号网络，与动物信号系统有重要的相关性。此外，ETMB调控的途径成分的发现可能为生物技术在重要农艺作物中的应用提供工具。本计划亦会为本科生和博士后提供跨学科研究的培训，包括招收少数族裔本科生，以论文、研讨会和会议报告的形式广泛传播研究成果，以及开设有关转译后修饰在信号通路中的作用的短期课程。

Various environmental and developmental stimuli generate reactive electrophile species (RES) that modify proteins through covalent adduct formation and trigger adaptive cellular responses in animal systems. Although this post-translational modification termed electrophile mediated thiol-binding (EMTB) is known to regulate signal transduction pathways in animals, it has not been previously reported in plant systems. Recently it was discovered that EMTB modification of a key enzyme in the oxylipin pathway of plants functions as an important signal in plant stress responses. The goal of this project is to examine the general role of EMTB modification in controlling plant signaling pathways.Intellectual Merit: Interdisciplinary approaches will be used to identify, on a genome-wide scale, the cellular target proteins that are covalently modified by stress-induced 12-oxophytodienoic acid (12-OPDA), a lipid RES in plants. A subset of the 12-OPDA binding protein targets will be fully characterized and examined for their role in plant stress signaling pathways. Finally, the specificity of 12-OPDA versus the other stress-induced thiolreactive electrophiles in covalent modification of the proteins identified will be determined. Broader impacts: The results of this study will provide important foundational insights into a novel, previously unrecognized posttranslational regulatory mechanism controlling a complex and crucial signaling network in plants with significant relevance to animal signaling systems. In addition the discovery of pathway components regulated by ETMB may provide for the generation of tools for biotechnological applications in agronomically important crops. This project will also provide for the training of undergraduate students and postdoctoral fellows in state-of-the-art interdisciplinary research and will include efforts to recruit underrepresented minority undergraduate students, the broad dissemination of research results in papers, seminars, and conference presentations, and the development of short courses on the role of post-translational modification in signaling pathways.