Post-translational Modifications as Modulators of Crop Plant Defense Signaling: a Systems Approach

作为作物防御信号调节剂的翻译后修饰：系统方法

• 批准号: 1238201
• 负责人: Jean Greenberg
• 金额: $ 135.92万
• 依托单位: University of Chicago
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-12-01 至 2017-11-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1238201&HistoricalAwards=false
• 关键词: Post; translational; Modifications; Modulators; Crop

PI: Jean T. Greenberg (University of Chicago)CoPIs: Stephen J. Kron (University of Chicago) and Howard C. Hang (Rockefeller University)The interaction of pathogenic microbes with plants elicits signaling pathways that can dictate the outcome of infection. Post-translational modifications (PTMs) elaborated after microbial perception enable a flexible mechanism for plants to activate networks of signaling events that promote host resistance. However, proteins such as acetyltransferase (AC-TR) effectors that are injected into plants by microbes can modify host proteins, which are then affected in their signaling functions due to alterations in folding, cellular location, enzymatic activity, ability to bind other proteins and/or stability. The recent availability of genomic sequence data of economically important plants and new technologies now makes it feasible to do systems-level global analysis of PTMs and to further dissect their impact on disease resistance. The goal of this project is to dissect how PTMs of plant proteins during infection modulates the ability of crop plants to defend against pathogenic bacteria. The project will (1) optimize in vitro methods for studying protein acetylation using AC-TR effectors from bacteria and host substrates; (2) perform global analyses of host protein acetylation in tomato using AC-TR effectors and test the role of select acetylated targets in plant defense; and (3) evaluate the impact of acetylation on defense signaling by analyzing global changes host protein phosphorylation, another PTM that is potentially competitive with acetylation. The project will develop reagents that will be used (and disseminated to the community) to quantitatively study signaling dynamics and relate changes in dynamics due to acetylation to infection outcomes. The global analysis of PTMs performed during this research will enhance the value of genomic studies and give a systems-level understanding of defense signaling that will expand the opportunities for crop improvements. The project will integrate research and teaching to provide interdisciplinary training to project personnel in proteomics, chemical biology, biochemistry, molecular genetics and plant-microbe interactions. It will also have broad impact by providing new tools and reagents to the community for analyzing defense signaling, and giving access to students from other universities to practical training through workshops on the analysis of PTMs. Inclusion of students from underrepresented groups will also have an impact on broadening participation in research of diverse groups. A major innovation of the project is the development and application of new methods for analyzing different acetylation sites in proteins at a global scale in relation to other potentially competitive modifications. The approach will be transformative for the study of acetylation in relation to other protein modifications in crop plants and applicable to a broad variety of plants and other organisms. A key outreach component will be three-day workshops for students from other institutions to learn the principles of detecting modifications of plant proteins. Students will gain hands-on experience on the generation and analysis of data on post-translational modifications using mass spectrometry. Additional outreach efforts will include participation in discussions about this project and demonstrations with school groups that visit the University of Chicago and Rockefeller University, coordination with teachers to modernize science curricula and discussions about research with the public at community fairs. This project will provide important interdisciplinary training in chemical biology, proteomics, biochemistry, molecular genetics and plant pathology to project personnel, including postdoctoral scholars and students at the graduate, post-baccalaureate, undergraduate and high school levels. Project personnel will be drawn from diverse groups to broaden participation in research science. Finally, the project will develop useful reagents (antibodies) and protocols to detect modifications in tomato proteins. The public will have access to the information generated by the project such as specific proteins that are subject to modification and approaches to monitor modifications through peer-reviewed publications and the project website. The public will also have access to reagents generated such as antibodies and tomato seeds as well as primary MS/MS spectra upon request. Proteomics datasets will also be deposited in one or more databases such as P3DB (http://p3db.org/).

PI：Jean T.格林伯格（芝加哥大学）CoPI：斯蒂芬J.克朗（芝加哥大学）和霍华德C。Hang（洛克菲勒大学）病原微生物与植物的相互作用激发了可以决定感染结果的信号通路。微生物感知后的翻译后修饰（PTM）使植物能够灵活地激活促进宿主抗性的信号事件网络。然而，通过微生物注射到植物中的蛋白质如乙酰转移酶（AC-TR）效应物可以修饰宿主蛋白质，然后由于折叠、细胞位置、酶活性、结合其他蛋白质的能力和/或稳定性的改变，宿主蛋白质的信号传导功能受到影响。最近获得的具有重要经济意义的植物和新技术的基因组序列数据，现在使得对PTM进行系统级全球分析并进一步剖析其对抗病性的影响成为可能。该项目的目标是剖析植物蛋白质在感染过程中的PTM如何调节作物抵抗病原菌的能力。该项目将（1）优化使用来自细菌和宿主底物的AC-TR效应子研究蛋白质乙酰化的体外方法;（2）使用AC-TR效应子对番茄中的宿主蛋白质乙酰化进行全面分析，并测试选择的乙酰化靶标在植物防御中的作用;和（3）通过分析宿主蛋白磷酸化的整体变化来评估乙酰化对防御信号的影响，另一种可能与乙酰化竞争的PTM。该项目将开发试剂，用于定量研究信号动力学，并将乙酰化引起的动力学变化与感染结果联系起来。在本研究期间进行的PTM全球分析将提高基因组研究的价值，并提供对防御信号的系统级理解，这将扩大作物改良的机会。该项目将结合研究和教学，为项目人员提供蛋白质组学、化学生物学、生物化学、分子遗传学和植物-微生物相互作用等方面的跨学科培训。它还将通过为社区提供新的工具和试剂来分析国防信号，并通过PTM分析研讨会为其他大学的学生提供实践培训，从而产生广泛的影响。吸收代表性不足群体的学生也将对扩大不同群体参与研究产生影响。该项目的一个主要创新是开发和应用新方法，用于在全球范围内分析蛋白质中与其他潜在竞争性修饰相关的不同乙酰化位点。该方法将是革命性的乙酰化与作物中其他蛋白质修饰的研究，并适用于各种植物和其他生物。一个关键的外联部分将是为来自其他机构的学生举办为期三天的讲习班，学习检测植物蛋白质修饰的原理。学生将获得使用质谱法生成和分析翻译后修饰数据的实践经验。其他外联工作将包括参与有关该项目的讨论，与访问芝加哥大学和洛克菲勒大学的学校团体进行示范，与教师协调使科学课程现代化，并在社区博览会上与公众讨论研究问题。该项目将为项目人员，包括博士后学者和研究生、学士后、本科和高中学生提供化学生物学、蛋白质组学、生物化学、分子遗传学和植物病理学方面的重要跨学科培训。项目人员将来自不同群体，以扩大对科学研究的参与。最后，该项目将开发有用的试剂（抗体）和检测番茄蛋白质修饰的方案。公众将可以通过同行评审的出版物和项目网站获得项目产生的信息，例如需要修改的特定蛋白质和监测修改的方法。 公众还可以根据要求获得生成的试剂，如抗体和番茄种子以及初级MS/MS光谱。蛋白质组学数据集也将被保存在一个或多个数据库中，例如P3 DB（http：//p3db.org/）。