Arabidopsis 2010: Analysis of Two-Component Signaling Elements from Arabidopsis

拟南芥 2010：拟南芥双组分信号元件分析

• 批准号: 0114965
• 负责人: Joseph Kieber
• 金额: $ 210万
• 依托单位: University of North Carolina at Chapel Hill
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0114965&HistoricalAwards=false
• 关键词: Arabidopsis; 2010; Analysis; Two; Component

Two-component systems are the primary means by which bacteria sense and respond to environmental stimuli. These systems are comprised of a number of distinct elements, namely histidine kinases, response regulators and in the case of phosphorelays, histidine phosphotransfer proteins (HPts). Genes encoding similar proteins to each of these elements have been identified in Arabidopsis, and for the majority of the 35 such genes no function has yet been definitively ascribed. An integrated approach to elucidate the function of these proteins in Arabidopsis is proposed. A combination of gene knockouts and inducible overexpression will be used to assess the roles of these genes in plant growth and development. The mutant plant lines will be characterized in terms of their response to biotic and abiotic factors such as hormones, light, and osmotic stress, and for their pattern of gene expression. Where in the plants these genes are expressed will be determined using a combination of GUS fusions and in situ RNA analysis. The location of the cognate proteins within the cell will also be delineated. To facilitate this localization, a series of 10 monoclonal antibodies will be generated to marker proteins, each of which resides on a distinct membrane. Protein complexes from Arabidopsis will be purified and analyzed to determine the interactions among these elements and to identify novel interacting proteins. Together, these studies will illuminate the signaling pathways in which each of these Arabidopsis two-component signaling elements function and how they interact to control plant growth and development.The data from these studies will be deposited on a publicly accessible web page that is currently under construction at UNC (http://www.bio.unc.edu/research/two-component/). A link to this web site will be established on the TAIR database, and we will coordinate with TAIR to deposit data as appropriate. The knockout seeds will be made publicly available through deposition in the ABRC Stock Center at Ohio State. The monoclonals raised against the membrane marker proteins will be available for the cost of shipping through UNH and the cell lines will also be deposited with the American Type Culture Collection (ATCC). In keeping with the goals of the 2010 project, functional analysis of the two-component signaling elements will aid in our understanding of plants at the organismal, cellular, and evolutionary levels. The research will provide functional information on the role of Arabidopsis two-component signaling systems in plant growth and development. The research will serve to define the subcellular location of the two-component signal transduction pathways, the interactions between the two-component signaling elements, and the downstream targets of the pathways. The research should clarify how a signal transduction mechanism that arose in bacteria has been adapted to plant signal transduction.These studies should uncover the functions of several gene families in Arabidopsis. The proteins encoded by these gene families are predicted to interact and thus our studies should aid in the development of a paradigm for signaling specificity among interacting members of large gene families. In addition, tools will be developed that will be generally applicable in defining the subcellular location of proteins in Arabidopsis.

两个组分系统是细菌感并响应环境刺激的主要手段。这些系统由许多不同的元素，即组氨酸激酶，反应调节剂，在磷酸层，组氨酸磷酸转移蛋白（HPTS）中。编码与这些元素中的每个元素相似的蛋白质的基因已在拟南芥中鉴定出来，并且对于35个这样的基因中的大多数，尚未确定尚无函数。提出了一种阐明这些蛋白质在拟南芥中的功能的综合方法。基因敲除和诱导过表达的结合将用于评估这些基因在植物生长和发育中的作用。突变植物系将以它们对生物和非生物因素的反应（例如激素，光和渗透胁迫）及其基因表达模式来表征。在植物中，这些基因表达的地方将使用GUS融合和原位RNA分析的组合确定。同源蛋白在细胞内的位置也将被描述。为了促进这种定位，将产生一系列10种单克隆抗体，以标记蛋白，每种蛋白都存在于不同的膜上。将纯化和分析来自拟南芥的蛋白质复合物，以确定这些元素之间的相互作用并鉴定新型相互作用的蛋白质。这些研究将共同​​阐明这些拟南芥两组式信号传导元素功能以及它们如何相互作用以控制植物生长和开发的信号通路。这些研究的数据将存放在目前正在UNC上的公共访问网页上，该网页正在UNC（http://wwwwwww.bio.unc.unc.unc.unc.unc.unc.unc.unc.unc.unceach/research/two-compention/two）中。将在TAIR数据库上建立指向此网站的链接，我们将与Tair协调以适当存放数据。淘汰种子将通过俄亥俄州立大学ABRC股票中心的沉积公开提供。针对膜标记蛋白提出的单克隆人将用于通过UNH的运输成本，并且细胞系也将与American Type Culture Collemon（ATCC）一起存放。为了与2010年项目的目标保持一致，对两组分的信号传导元件的功能分析将有助于我们对生物，细胞和进化水平的植物的理解。这项研究将提供有关拟南芥两组分信号系统在植物生长和发育中的作用的功能信息。 该研究将用于定义两个组分信号转导途径的亚细胞位置，两组分量信号传导元件之间的相互作用以及途径的下游目标。 该研究应阐明细菌中产生的信号转导机制如何适应植物信号转导。这些研究应揭示拟南芥中几个基因家族的功能。 这些基因家族编码的蛋白质被预计会相互作用，因此我们的研究应有助于开发大基因家族相互作用成员之间信号特异性的范式。 此外，将开发工具通常适用于定义拟南芥中蛋白质的亚细胞位置。