Proteomics analysis of endosomal compartments in Arabidopsis (ERA-PG 069)

该提案涉及“有助于欧洲植物基因组学研究领域发展的其他主题”（子呼叫A）。包括产量稳定性在内的植物性能在很大程度上取决于与生物和非生物环境的相互作用以及植物本身的细胞间通讯。营养摄取和信号传导是由质膜定位的蛋白质介导的，如转运蛋白和受体，其活性取决于与下游效应物的相互作用，以及它们通过内体运输和分类的可用性。内体分选的结果要么是通过靶向液泡降解蛋白质，要么是蛋白质再循环到质膜。本项目旨在利用蛋白质组学方法对拟南芥的内体区室进行分析，为植物性能的功能研究提供必要的基础。具体来说，免疫分离的、超微结构定义的内体区室的驻留蛋白和货物蛋白将通过质谱鉴定，随后在表达myc标记或gfp融合版本的新鉴定蛋白的转基因植物中分析内体定位。预期的结果将确定内体室在质膜蛋白运输中的作用，并使早期/分类和回收内体得以区分。

This proposal addresses "other topics that contribute to the development of the European Research Area in plant genomics" (Sub Call A). Plant performance including yield stability largely depends on the interaction with the biotic and abiotic environment as well as on cell-cell communication within the plant itself. Nutrient uptake and signaling are mediated by plasma membrane-localised proteins such as transporters and receptors whose activity is determined by interaction with downstream effectors as well as their availability through endosomal trafficking and sorting. Endosomal sorting results either in protein degradation via targeting to the vacuole or in protein recycling to the plasma membrane. To provide a necessary foundation for functional studies of plant performance as specified in other topics of Sub Call A, this project aims to analyse endosomal compartments of Arabidopsis by proteomics. Specifically, resident and cargo proteins of immuno-isolated, ultrastructurally defined endosomal compartments will be identified by MS and subsequently analysed for endosomal localisation in transgenic plants expressing Myc-tagged or GFP-fusion versions of newly-identified proteins. The anticipated results will establish the role of endosomal compartments in plasma-membrane protein trafficking and enable distinction of early/sorting from recycling endosomes.