Synaptotagmin A and C in virus movement and plant development

突触结合蛋白 A 和 C 在病毒运动和植物发育中的作用

基本信息

批准号：
8109984
负责人：
Asako Uchiyama
金额：
$ 5.22万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2012-09-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8109984
关键词：
Agriculture Alleles Animal Model Animals Arabidopsis Begomoviruses Binding Biolistics Biological Assay Cabbage - dietary Cell Wall Cell membrane Cell physiology Cells Chromosome Mapping Complex Complication Confocal Microscopy Development Early Endosome Endocytosis Engineering Exocytosis Geminiviridae Gene Family Gene Fusion Genes Genetic Genome Genomics Goals HIV Human Human Virus In Vitro Invaded Invertebrates Investigation Lead Left Life Macromolecular Complexes Mediating Membrane Modeling Molecular Mechanisms of Action Mouse-ear Cress Movement Mutate Neurons Organism Pathway interactions Pattern Phenotype Plant Leaves Plant Model Plant Viruses Plants Plasmodesmata Play Polymorphic Microsatellite Marker Process Production Property Protein Binding Proteins Reporter Genes Reporting Research Resistance Resources Retroviridae Role Seeds Son of Sevenless Proteins Structure Synaptic Vesicles System T-DNA Testing Time Tobacco Mosaic Virus Tobamovirus Transgenic Organisms Vesicle Viral Viral Genome Virus Virus Assembly Yeasts base cell motility cellular imaging improved insight mutant neurotransmitter release nutrition plant growth/development promoter protein function protein transport relating to nervous system sensor synaptotagmin trafficking vector

项目摘要

DESCRIPTION (provided by applicant): Synaptotagmins are multidomain proteins belonging to a large gene family in animals, which are exclusively or predominantly expressed in neuronal cells. They are proposed to be the Ca2+ sensors that trigger rapid and synchronous synaptic vesicle exocytosis to release neurotransmitter, and subsequent endocytosis to recapture membrane. However, the functions of synaptotgamins in non-neural cells have not been well characterized. Plant virus cell-to-cell spread through the barrier of the plant cell wall is mediated by virus-encoded movement proteins (MPs). Recently, the investigation of this process lead to the unexpected discovery of a five-gene family of synaptotagmins (AtSYTA through AtSYTE) in the model plant Arabidopsis thaliana. Our lab has shown that: (1) SYTA interacts with, and regulates the cell-to-cell trafficking of, the distinct movement proteins (MPs) encoded by a Begomovirus and a Tobamovirus; and (2) SYTA regulates the formation of early endosomes at the plasma membrane in plant cells. We propose that SYTA regulates plant virus cell-to-cell movement, and plant cell macromolecular trafficking, through, plasmodesmata via an endocytic recapture pathway. Our long term goal is to understand the roles of Syts in plant cell function and plant development, and to understand how viruses usurp the endocytic machinery to exit from an infected cell. The proposed research will focus on characterizing the interactions of SYTA with a variety of diverse virus movement proteins, and the functions of SYTC in virus movement and plant development. Two specific aims are proposed: (1) To investigate the roles of SYTA in virus movement using cell trafficking, infectivity, in vitro protein binding, and live, cell imaging assays; and (2) To determine if SYTA and SYTC have overlapping functions in virus movement and in plant development by characterizing sytc and syta/sytc mutant Arabidopsis lines, determining developmental pattern of SYTC expression, and characterizing the interaction of SYTC with viral MPs. This research will utilize a newly developed biolistic assay to transiently express proteins in Arabidopsis leaf cells and multiphoton confocal microscopy to examine protein interactions in intact leaves in real time. The proposed research can lead to rational approaches to engineer virus-resistant plants, thus improving agricultural production and human nutrition. Beyond this, these studies can inform research in invertebrate and mammalian systems on the functions of Syts in non-neural cells by providing new insights relevant to understanding the roles of animal Syts and vesicle trafficking in development, and how animal and human viruses such as retroviruses like HIV may take advantage of vesicle trafficking for virus assembly and exit.

描述（由申请人提供）：突触毒素是属于动物中大基因家族的多域蛋白，这些蛋白是在神经元细胞中仅或主要表达的。认为它们是触发快速和同步突触囊泡外胞毒性释放神经递质的CA2+传感器，随后的内吞作用释放了膜。然而，非神经细胞中突触毒素的功能尚未得到很好的特征。通过病毒编码的运动蛋白（MP）介导植物病毒细胞到细胞通过植物细胞壁的屏障传播。最近，对这一过程的调查导致在模型植物拟南芥中意外地发现了一个五基因的突触毒素家族（Atsyta通过Atsyte）。我们的实验室表明：（1）SYTA与由Begomovirus和Tobamovirus编码的独特运动蛋白（MPS）相互作用并调节细胞对细胞运输；（2）SYTA调节植物细胞中质膜早期内体的形成。我们建议SYTA通过内吞重复途径来调节植物病毒细胞向细胞运动，并通过质量分子的植物细胞大分子运输。我们的长期目标是了解SYT在植物细胞功能和植物发育中的作用，并了解病毒如何使内吞机械从受感染的细胞中退出。拟议的研究将着重于表征SYTA与多种不同病毒运动蛋白的相互作用，以及SYTC在病毒运动和植物发育中的功能。提出了两个具体目的：（1）使用细胞运输，感染性，体外蛋白质结合和活细胞成像测定法研究SYTA在病毒运动中的作用；（2）确定SYTA和SYTC是否通过表征SYTC和SYTA/SYTA/SYTC突变拟南芥线条来确定SYTC表达的发育模式以及表征SYTC与病毒MPS的相互作用来确定SYTA和SYTA/SYTC突变拟南芥线的重叠函数。这项研究将利用新开发的生物学测定法在拟南芥叶细胞和多极子共聚焦显微镜中瞬时表达蛋白质，以实时检查完整叶片中的蛋白质相互作用。拟议的研究可以导致对耐药植物的合理方法，从而改善农业生产和人类营养。除此之外，这些研究还可以通过提供与了解动物Syts和囊泡运输在发育中的动物和囊泡贩运在发育中的作用以及诸如HIV之类的动物和人类病毒（如HIV）等新见解的新见解，从而为非神经细胞中SYT的功能提供了有关SYT在非神经细胞中的功能的研究的信息，例如HIV可能会利用Virus的速度和Exit。