Understanding the role of the plastid outer envelope membrane for integrating plastids into cellular metabolic and regulatory networks

这个合作研究项目的目的是确定叶绿体外被膜在整合质体与植物细胞中的代谢和调节网络中的作用。为此，定位于豌豆的外质体包膜膜的蛋白质将通过蛋白质组学鉴定。它们在植物细胞中的定位将在拟南芥中通过用绿色荧光蛋白标记和双砷染料测定来独立评估。将对豌豆进行全面的并行转录组测序，以生成蛋白质组学所需的序列数据库。将使用由NSF Arabidopsis 2010计划资助在密歇根州立大学建立的突变体和数据分析管道，分离并分析质体外被膜蛋白质编码基因缺陷的拟南芥突变体，以及其他数千种质体靶向和过氧化物酶体蛋白质缺陷的突变体。通过这个AFGN项目资助的研究生将在密歇根州立大学接受大规模突变和多变量数据分析的培训。该项目将为植物研究界提供细胞内蛋白质定位的信息和豌豆转录组的综合数据库。使用一个一致的管道数千突变体的分析将提供一个独特的机会，深入了解质体包膜膜的作用，在整合质体与其他细胞室，并产生可检验的蛋白质功能的假设。

This collaborative research project aims at identifying the role of the chloroplast outer envelope membrane in integrating plastids with the metabolic and regulatory networks in plant cells. To this end, proteins localized in the outer plastid envelope membrane of pea will be identified by proteomics. Their localization in plant cells will be independently assessed in Arabidopsis by tagging with the green fluorescent protein and biarsenical-dye assays. Comprehensive massively-parallel transcriptome sequencing of pea will be conducted to generate a sequence database that is required for proteomics. Arabidopsis mutants deficient in genes encoding proteins of the plastid outer envelope membrane will be isolated and analyzed jointly with several thousand other mutants deficient in plastid-targeted and peroxisomal proteins, using a mutant and data analysis pipeline that was established at Michigan State University with funding from the NSF Arabidopsis 2010 program. Graduate students funded through this AFGN project will be trained in large-scale mutant and multivariate data analysis at Michigan State University. This project will provide the plant research community with information on intracellular protein localization and with a comprehensive database of the pea transcriptome. Using a consistent pipeline for the analysis of several thousand mutants will provide a unique opportunity to gain insight into the role of the plastid envelope membrane in integrating plastids with other cellular compartments and for generating testable hypotheses of protein function.