Molecular Analysis of Proteins Involved in Plant Endocytosis and Cytokinesis

正常的植物生长和发育高度依赖于网格蛋白介导的膜运输，以进行细胞分裂、对生长素运输、营养吸收、激素和病原体信号传递至关重要的质膜蛋白的摄取和再循环。十多年的研究使我们对酵母和哺乳动物细胞内吞作用所需的蛋白质和脂质复杂网络有了目前的了解。相比之下，我们对植物中网格蛋白介导的膜运输机制的了解还很初级。然而，最近的研究表明，植物的内吞机制及其调控已经进化到满足植物形态发生和生长所必需的独特要求。了解植物中网格蛋白介导的膜转运与其他系统之间的异同的关键是全面鉴定和表征进化上保守的，以及涉及植物网格蛋白包被囊泡形成和靶向的植物特异性核心和辅助机制，并阐明这些蛋白质在植物细胞分裂中的作用。通过蛋白质组学、生物化学、遗传学和影像学研究来鉴定和直接表征参与植物网格蛋白包被的内吞囊泡形成和靶向的蛋白质。这些研究可能会为植物和进化分化生物中依赖网格蛋白的贩运的分子动力学提供新的见解。此外，由于网格蛋白包被囊泡运输涉及广泛的植物过程，我们的研究结果将产生基础知识，从长远来看，将有助于开发具有可持续粮食、饲料和生物燃料生产所必需的生物质产量的改良作物。这项工作还将涉及本科生、研究生和博士后的培训，他们将学习各种重要的技术，包括成像的计算技术。

Intellectual MeritNormal plant growth and development is highly dependent upon clathrin-mediated membrane trafficking for cell division, the uptake and recycling of plasma membrane proteins critical for auxin transport, nutrient uptake and hormone and pathogen signaling. More than a decade of research has led to our current understanding of the complex network of proteins and lipids required for endocytosis in yeast and mammalian cells. By comparison, our knowledge of the mechanism of clathrin-mediated membrane transport in plants is rudimentary. However, recent studies have demonstrated that the plant endocytic machinery and its regulation have evolved to meet the unique requirements necessary for plant morphogenesis and growth. Key to our understanding the similarities and differences between clathrin-mediated membrane transport in plants and other systems will be the comprehensive identification and characterization of the evolutionarily conserved, along with the plant-specific core and accessory machinery involved in the formation and targeting of plant clathrin-coated vesicles and elucidation of the role of these proteins in plant cell division. Proteomic, biochemical, genetic and imaging studies are proposed to identify and directly characterize proteins involved in the formation and targeting of plant clathrin-coated endocytic vesicles. Broader Impact These studies will likely provide new insight into the molecular dynamics of clathrin-dependent trafficking in plants and evolutionarily divergent organisms. In addition, as clathrin-coated vesicle trafficking is involved in a wide range of plant processes, our results should generate fundamental knowledge that will in the long term, aid in the development of improved crops with enhanced biomass yield necessary for sustainable food, feed, and biofuel production. The work will also involve the training of undergraduates, graduate students and post-docs who will learn a variety of important techniques including computational techniques for imaging.