An Interdisciplinary Approach to Deciphering the Molecular Dialogue between the Plasma Membrane and Nucleus of Medicago truncatula

项目负责人：Michael R. Sussman（威斯康星大学麦迪逊分校）项目负责人：Jean Michel an<s:1>、Joshua J. Coon和Sushmita Roy（威斯康星大学麦迪逊分校）该项目的目标是开发和应用基因组技术，以在分子水平上理解细菌和植物如何交流，从而创造出一种非凡的进化产物，即固氮共生结核。微生物和植物之间的这种对话的一个重要部分是细胞内蛋白质化学组成的变化，即磷酸基团在特定氨基酸上添加或去除的程度。使用最先进的质谱仪来精确确定哪些蛋白质被磷酸盐修饰，以及细菌与植物相遇的第一个小时内的蛋白质修饰量，该项目将试图阐明植物如何改变其蛋白质，以启动它们的代谢和基因表达的无数变化，这些变化是共生结节形成所必需的。所有项目成果（如数据、多肽和种质资源）将免费向公众提供，不受知识产权限制。在这个项目中产生的Medicago转录组、蛋白质组和磷蛋白质组数据将存储在国家生物技术信息中心（NCBI: http://www.ncbi.nlm.nih.gov/）、豆类信息服务（LIS: http://www.comparative-legumes.org/）和肽图谱（www.peptideatlas.org）等长期存储库中，并向公众开放。在这个项目中开发和使用的质谱技术可以用于了解许多不同的生物事件，除了共生固氮。为了给美国各地的研究人员提供机会，使他们能够使用这种先进的仪器和技术进行研究，该项目将开发并举办为期一周的植物蛋白质组学研讨会。这个研讨会每年夏天在威斯康星大学举行，将为20多名参与者（研究生、博士后研究人员和教授）提供动手的机会，深入使用质谱仪在他们自己的实验室里测量正在研究的植物蛋白质的变化。这次研讨会将极大地促进这项技术在植物生物学一般领域的研究的更广泛的可及性和使用，并将极大地帮助我国培养我们未来的植物科学家的目标，并帮助他们适应现代基因组“系统生物学”实验技术。

PI: Michael R. Sussman (University of Wisconsin - Madison)CoPIs: Jean Michel Ané, Joshua J. Coon, and Sushmita Roy (University of Wisconsin - Madison)This goal of this project is to develop and apply genomic technologies for understanding at a molecular level, how bacteria and plants communicate to create a remarkable product of evolution, the nitrogen-fixing symbiotic nodule. An important part of this conversation between the microbe and the plant is a change in the chemical composition of proteins within the cell, i.e., in the degree by which phosphate groups are added or removed to specific amino acids. Using state of the art mass spectrometers to determine precisely which proteins are being modified by phosphate, and by how much in the first hour in which the bacteria meets the plant, the project will attempt to elucidate how the plant changes its proteins in order to initiate all of the myriad changes in their metabolism and gene expression that are required for the symbiotic nodules to form. All project outcomes (e.g., data, peptides and germplasm resources) will be provided to the public free of charge and without intellectual property restrictions. Medicago transcriptome, proteome and phosphoproteome data generated in this project will be deposited and accessible to the public at long-term repositories such as National Center for Biotechnology Information (NCBI: http://www.ncbi.nlm.nih.gov/), the Legume Information Service (LIS: http://www.comparative-legumes.org/) and the PeptideAtlas (www.peptideatlas.org). The mass spectrometric technology being developed and used in this project could be useful for understanding many different biological events, besides symbiotic nitrogen fixation. In order to provide opportunities for researchers all over the U.S. to access this sophisticated instrumentation and technology for use in their research, the project will develop and hold a one-week long Plant Proteomic Workshop. To be held each summer at the University of Wisconsin, this workshop will provide two dozen participants (graduate students, postdoctoral researchers and professors) with hands-on, in-depth use of mass spectrometers for measuring changes in the proteins from plants under study in their own labs. This workshop will greatly facilitate the more widespread accessibility and use of this technology for research in the general area of plant biology, and should greatly help this country's goal of training our future plant scientists and help them become comfortable with modern genomic "systems biology" experimental techniques.