Synaptotagmin function in virus movement and plant development

突触结合蛋白在病毒运动和植物发育中的功能

• 批准号: 7199365
• 负责人: SONDRA G LAZAROWITZ
• 金额: $ 36.03万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7199365
• 关键词: Address; Animals; Arabidopsis; Arabidopsis Proteins; Begomoviruses; Binding; Biochemical; Biological Assay; Brain; Cabbage - dietary; Calcium; Capsid Proteins; Cell Communication; Cell Wall; Cell physiology; Cells; Complex; Defect; Development; Disease Resistance; Early Endosome; Endocytosis; Endosomes; Engineering; Exocytosis; FM 4-64; Funding; Gene Family; Gene Proteins; Genes; Goals; Growth; HIV; Health; Human; Image; Immune; In Vitro; Individual; Insertional Mutagenesis; Invaded; Knowledge; Label; Left; Life; Link; Mouse-ear Cress; Movement; Multigene Family; Mutate; Names; Neuroendocrine Cell; Nuclear; Orthologous Gene; PPBP gene; Pathway interactions; Plant Leaves; Plant Model; Plant Viruses; Plants; Plasmodesmata; Play; Process; Proteins; Public Health; RNA Interference; Reading; Recycling; Regulation; Research; Research Personnel; Retroviridae; Role; Signal Transduction; Son of Sevenless Proteins; Squash; Synaptic Vesicles; T-DNA; Testing; Thinking; Time; Tobacco Mosaic Virus; Tobamovirus; Transgenic Organisms; Transgenic Plants; Vesicle; Viral; Viral Genome; Virus; Yeasts; base; cell motility; cellular imaging; disorder control; improved; insight; neurotransmitter release; novel; nutrition; plant growth/development; plant virus movement protein; programs; promoter; protein transport; sensor; synaptotagmin; trafficking

DESCRIPTION (provided by applicant): Plant virus movement proteins transport viral genomes across the cell wall by altering plasmodesmata, transwall pores that connect adjacent plant cells. Synaptotagmins, a multigene family thought to be exclusive to animals, are calcium (Ca2+) sensors that regulate synaptic vesicle exo/endocytosis. Our long term goal is to understand how synaptotagmins regulate virus movement and intercellular transport to control disease resistance, the spread of RNAi signals and cell fate. SYTA is 1 of 5 Arabidopsis synaptotagmins (SYT A-E). SYTA binds to the distinct movement proteins encoded by the Begomoviruses CaLCuV (MPCaLCuV) and SqLCV, and the Tobamovirus TMV (30K). We have shown that SYTA regulates both the formation of early endosomes and the cell-to-cell transport of MPCaLCuV and TMV 30K. We propose that SYTs regulate macromolecular trafficking to plasmodesmata via an endocytic recapture pathway, and potentially entry into and exit from the pore as well. We will use a combination of transgenic plant studies and cell-based expression assays to investigate if SYTs A, C and E are key regulators of plant virus movement and intercellular transport. Biochemical and infectivity studies, and a leaf-based cell trafficking assay will examine if SYTA alone regulates the cell-to-cell movement of most viruses, or whether SYTC and SYTE are also involved, either independent of or partnering with SYTA. The roles of SYTC and SYTE in vesicle trafficking will be defined by immune localization, and the use of cell- and leaf-based secretion assays. Multiphoton imaging of SYTA itself, or with MPCaLCuV and 30K, or SYTC and SYTE, will show how SYTA endosomes ferry movement proteins to plasmodesmata, and how SYTC or SYTE may cooperate in this process. This knowledge of SYT A, C and E action in plant cells will be connected to the regulation of development through promoter::GUS fusion studies to define their spatial and temporal expression, and analysis of transgenic lines in which the expression of each is inhibited by RNAi or T-DNA insertions. Our demonstration of a SYTA-regulated functional link between early endosome formation and intercellular transport provides novel insights into how plasmodesmal function is controlled. Beyond the agronomic impact of engineering disease-resistant plants, our studies are relevant to human health in that they can inform and accelerate studies of animal synaptotagmins in cell-cell communication and development, and of mechanisms by which retroviruses redirect the endosomal machinery for virus maturation and exit from an infected cell. This research investigates the regulation of what seems to be a common pathway for transport between plant and animal cells. As such, it can impact public health by improving nutrition, providing knowledge about growth defects, and identifying novel ways to limit the spread of HIV and other retroviruses.

描述（由申请人提供）：植物病毒运动蛋白通过改变胞间连丝、连接相邻植物细胞的穿壁孔，将病毒基因组运输穿过细胞壁。突触结合蛋白是一个被认为是动物独有的多基因家族，是调节突触小泡胞吐/内吞作用的钙 (Ca2+) 传感器。我们的长期目标是了解突触结合蛋白如何调节病毒运动和细胞间运输，以控制疾病抵抗力、RNAi 信号的传播和细胞命运。 SYTA 是 5 种拟南芥突触结合蛋白 (SYTA-E) 中的一种。 SYTA 与 Begomoviruses CaLCuV (MPCaLCuV) 和 SqLCV 以及 Tobamovirus TMV (30K) 编码的不同运动蛋白结合。我们已经证明 SYTA 调节早期内体的形成以及 MPCaLCuV 和 TMV 30K 的细胞间转运。我们认为 SYT 通过内吞重捕获途径调节大分子向胞间连丝的运输，并可能进入和离开孔。我们将结合转基因植物研究和基于细胞的表达测定来研究 SYT A、C 和 E 是否是植物病毒运动和细胞间运输的关键调节因子。生化和感染性研究以及基于叶子的细胞运输测定将检查 SYTA 是否单独调节大多数病毒的细胞间运动，或者 SYTC 和 SYTE 是否也参与其中，无论是独立于 SYTA 还是与 SYTA 合作。 SYTC 和 SYTE 在囊泡运输中的作用将通过免疫定位以及基于细胞和叶子的分泌测定的使用来定义。 SYTA 本身的多光子成像，或使用 MPCaLCuV 和 30K，或 SYTC 和 SYTE，将显示 SYTA 内涵体如何将运动蛋白运送到胞间连丝，以及 SYTC 或 SYTE 如何在此过程中合作。对植物细胞中 SYT A、C 和 E 作用的了解将与通过启动子::GUS 融合研究来定义其空间和时间表达的发育调节以及转基因株系的分析（其中每个基因的表达受到 RNAi 或 T-DNA 插入的抑制）联系起来。我们对早期内体形成和细胞间运输之间 SYTA 调节的功能联系的论证为如何控制胞间连丝功能提供了新的见解。除了工程抗病植物的农艺影响之外，我们的研究还与人类健康相关，因为它们可以为细胞间通讯和发育中的动物突触结合蛋白以及逆转录病毒重定向内体机制以促进病毒成熟和从受感染细胞中退出的机制提供信息和加速研究。这项研究调查了植物和动物细胞之间似乎常见的运输途径的调节。因此，它可以通过改善营养、提供有关生长缺陷的知识以及确定限制艾滋病毒和其他逆转录病毒传播的新方法来影响公共健康。