Arabidopsis 2010 (Collaborative Project): Understanding Peroxisomal Protein Networks

植物过氧化物酶体是多种生理和代谢途径的重要组成部分，但我们对这些细胞器的蛋白质组成和代谢调控网络的了解还远远不够。这个2010年的项目将使用生物信息学和蛋白质组学，然后是荧光显微镜和蛋白质分离，生成过氧化物酶体基质和膜的“零件清单”。此外，过氧化物酶体靶向蛋白的功能将通过序列索引T-DNA插入突变体的各种检测来分析。最后，过氧化物酶体蛋白复合物和网络将探索，使用凝胶为基础的方法结合质谱。大约300-500个编码过氧化物酶体蛋白的基因将与生物学功能相关，这是建立植物过氧化物酶体功能综合模型的关键一步。该项目将为科学界提供急需的过氧化物酶体蛋白质的全面清单和大量标记的报告构建物，用于后续研究。所有信息将被整合到AraPeroX数据库（http://www.araperox.uni-goettingen.de/）中，并镜像到项目网站http://www.peroxisome.msu.edu上。生成的信息和材料将通过我们的项目网站、TAIR和ABRC及时向公众提供。考虑到不同的细胞区室通常通过过氧化物酶体功能连接，并且过氧化物酶体在逆境反应中至关重要，该项目将使植物群落的多个领域受益，并与2010年正在进行的其他项目（如质体功能基因组学项目）产生协同影响。这一研究结果也将扩大我们对真核细胞生物学的认识，并为工程作物提高抗逆性和提高产油量提供有用的信息。更广泛的影响：通过对本科生、高中生和教师以及主要本科院校的教师进行多学科培训，将研究和教育结合起来。项目中产生的数据、资源和技术将对广泛的科学家社区有用。

Plant peroxisomes are essential components in various physiological and metabolic pathways, yet our knowledge of the protein composition and metabolic and regulatory networks associated with these organelles is far from complete. This 2010 project will generate a 'parts list' of peroxisomal matrix and membrane, using bioinformatics and proteomics, followed by fluorescence microscopy and protein fractionation. Further, the functions of peroxisome-targeted proteins will be analyzed with various assays on sequence-indexed T-DNA insertion mutants. Finally, peroxisomal protein complexes and networks will be explored, using gel-based methods in combination with mass spectrometry. Approximately 300-500 genes encoding peroxisomal proteins will be associated with biological functions, which is a critical step toward developing a comprehensive model for plant peroxisome function. This project will provide the scientific community with a much-needed comprehensive inventory of peroxisomal proteins and a large set of tagged reporter constructs for follow-up studies. All information will be integrated into the AraPeroX database (http://www.araperox.uni-goettingen.de/) and mirrored on the project website http://www.peroxisome.msu.edu. Information and materials generated will be available to the public in a timely fashion through our project website, TAIR, and ABRC. Given that different cell compartments are often linked via peroxisomal functions and that peroxisomes are crucial during stress responses, this project will benefit several fields in the plant community and achieve synergistic impact with other ongoing 2010 projects, such as the plastid functional genomics project. Results from this research will also expand our knowledge of eukaryotic cell biology and provide useful information to engineering crop plants for increased stress resistance and improved oil production.Broader Impacts: Research and education will be integrated by multidisciplinary training of undergraduates, high school students and teachers, and teachers from primarily undergraduate colleges. The data, resources, and techniques generated in the project will be useful to a broad community of scientists.