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

• 批准号: 8132804
• 负责人: Asako Uchiyama
• 金额: $ 5.47万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 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/8132804
• 关键词: Agriculture; Alleles; Animal Model; Animal Viruses; 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; 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

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）：突触结合蛋白是属于动物中的大基因家族的多结构域蛋白，其仅在或主要在神经元细胞中表达。它们被认为是Ca 2+感受器，触发快速和同步的突触囊泡胞吐作用以释放神经递质，以及随后的胞吞作用以重新捕获膜。然而，突触结合素在非神经细胞中的功能还没有得到很好的表征。植物病毒通过植物细胞壁的屏障在细胞间传播是由病毒编码的运动蛋白（MP）介导的。最近，对这一过程的研究导致在模式植物拟南芥中意外地发现了一个由五个基因组成的突触结合蛋白家族（AtSYTA至AtSYTE）。我们的实验室已经证明：（1）SYTA与由菜豆病毒属和烟草花叶病毒属编码的不同运动蛋白（MP）相互作用，并调节其细胞间运输;和（2）SYTA调节植物细胞质膜上早期内体的形成。我们建议，SYTA调节植物病毒细胞间的运动，和植物细胞大分子运输，通过，胞间连丝通过内吞再捕获途径。我们的长期目标是了解Syts在植物细胞功能和植物发育中的作用，并了解病毒如何篡夺内吞机制从受感染的细胞中退出。该研究将集中在表征SYTA与各种不同的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。本研究的主要目的是：（1）利用细胞运输、感染性、体外蛋白结合和活细胞成像等方法研究SYTA在病毒运动中的作用;和（2）通过表征sytc和syta/sytc突变体拟南芥系，确定SYTC表达的发育模式，以及表征SYTC与病毒MP的相互作用。本研究将利用一种新开发的生物射弹技术在拟南芥叶细胞中瞬时表达蛋白质，并利用多光子共聚焦显微镜在真实的时间内研究完整叶片中蛋白质的相互作用。拟议的研究可以导致合理的方法来设计抗病毒植物，从而改善农业生产和人类营养。除此之外，这些研究可以通过提供与理解动物Syts和囊泡运输在发育中的作用相关的新见解，以及动物和人类病毒（如HIV等逆转录病毒）如何利用囊泡运输进行病毒组装和退出，为无脊椎动物和哺乳动物系统中Syts在非神经细胞中的功能提供信息。