Arabidopsis 2010: Protein Interacting Networks and Site-Specific Phosphorylation in Leucine-Rich Repeat Receptor-Like Kinase Function

植物和动物细胞通过受体感知和响应环境中的信号，这些受体通过蛋白质-蛋白质相互作用将生化信息传递到细胞核，在细胞核中发生基因表达的变化。其中一类受体（富含亮氨酸重复受体样激酶或LRR RLKs）存在于模式植物拟南芥中223个成员的大家族中。这些受体的结构组织与动物受体激酶相似，其中一些已被详细研究，是调节植物生长、形态发生、抗病和应激反应的关键因素。然而，绝大多数信号转导分子家族的功能仍然未知。在美国国家科学基金会拟南芥2010计划的前期支持下，研究人员已经生成了一套遗传和生化资源，将允许深入研究LRR RLKs在植物生长和胁迫反应中的功能。使用蛋白质组学和质谱方法，该项目旨在定义与这些LRR RLKs相互作用的蛋白质。这些蛋白质形成了一个相互作用的网络，其功能最终决定了一系列生理反应，本项目将定义这个网络的组成和性质。除了对蛋白质网络相互作用的广泛研究外，还将在详细的生化水平上对选定的一组LRR-RLKs进行表征，包括功能分析，从而了解这些蛋白质如何影响植物生长。研究的广泛影响：鉴于已知LRR RLK家族对植物发育的重要性，该项目的结果可能会对农业产生实际影响。此外，该项目将为拟南芥研究界提供广泛的资源，包括一个数据库（http://www4.ncsu.edu/~sclouse/Clouse2010.htm），该数据库为LRR RLK蛋白网络和磷酸化位点的比较研究创造了独特的资源。美国国家科学基金会支持的非常成功的植物磷酸化研讨会将重新启动，并将为多达45个实验室的研究人员提供互动和分享研究想法和结果的论坛。该项目还将提供从高中实习生到博士后科学家的各级蛋白质生物化学、质谱和拟南芥分子遗传学方面的培训。对这些人的招募将与北卡罗莱纳州立大学和伊利诺伊大学的既定项目相结合，这些项目鼓励未被充分代表的少数民族参与科学研究。

Plant and animal cells sense and respond to signals in their environment through receptors that relay biochemical information via protein-protein interactions to the nucleus, where changes in gene expression occur. One class of these receptors (the Leucine-Rich Repeat Receptor-like kinases or LRR RLKs) is present in a large family of 223 members in the model plant Arabidopsis thaliana. These receptors have a structural organization similar to that of animal receptor kinases, and several that have been studied in detail are known to be critical elements that regulate the plant's growth, morphogenesis, disease resistance, and responses to stress. However, the functions of the vast majority of this large family of signal transduction molecules remain unknown. With previous support from the NSF Arabidopsis 2010 Program, the investigators have generated a set of genetic and biochemical resources that will allow the in-depth study of the function of LRR RLKs in plant growth and stress responses. Using proteomic and mass spectrometry approaches, this project is designed to define the proteins that interact with these LRR RLKs. These proteins form an interactive network whose function ultimately determines a range of physiological responses, and this project will define the components and properties of this network. In addition to this broad study of protein network interactions, a selected group of LRR-RLKs will be characterized at a detailed biochemical level, including functional analyses that will permit an understanding of how these proteins influence plant growth.Broader Impact of the Research: Given the known importance of the LRR RLK family to plant development, the results of this project are likely to have practical agricultural impact. In addition this project will provide extensive resources to the Arabidopsis research community including a database (http://www4.ncsu.edu/~sclouse/Clouse2010.htm) that creates a unique resource for the comparative study of LRR RLK protein networks and phosphorylation sites. The highly successful NSF-supported Plant Phosphorylation Workshop will be reinitiated and will serve as a forum for researchers from as many as 45 laboratories to interact and share research ideas and results. The project will also provide training in protein biochemistry, mass spectrometry, and Arabidopsis molecular genetics at all levels from high school student interns through postdoctoral scientists. Recruitment of these individuals will be integrated with established programs at North Carolina State University and the University of Illinois that encourage the participation of under-represented minorities in scientific research.