2010/AFGN Collaborative Project: The Interaction Network for the Heterotrimeric G-Protein Signaling Pathway

2010/AFGN 合作项目：异三聚体 G 蛋白信号通路的相互作用网络

• 批准号: 49640339
• 负责人: Professor Dr. Ralph Panstruga
• 金额: --
• 依托单位: Department of Biology
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2007
• 资助国家: 德国
• 起止时间: 2006-12-31 至 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/49640339?language=en
• 关键词: 2010; AFGN; Collaborative; Project; Interaction

A variety of signaling networks in Arabidopsis utilize heterotrimeric G-protein complexes to couple signals that are perceived at the cell surface by G-protein coupled receptors (GPCRs) to enzymes called effectors. To date, neither the downstream effectors nor the upstream GPCRs have been unequivocally identified. This paucity of information stands in contrast to human and C. elegans G protein-coupled pathways where 950 GPCRs and more than a dozen effectors have been identified thus representing the most extensive signaling network in metazoans. We will use a high through-put interaction screen to identify proteins that interact with the heterotrimeric G protein complex. Interactors will be tested for in vivo interaction and confirmed candidates will be used as bait to initiate additional rounds of screening. The data will be subjected to bioinformatic and graph-theoretical meta-analyses to identify the main signal transduction pathways and protein complexes (signalosomes) in the interaction network. Selected interactors will be pursued more thoroughly. Reverse genetic and molecular biological approaches will be applied to determine on which branches of the G-protein signaling network these elements operate. Up to 100 genes encoding confirmed interactors will be knocked out either by insertional mutagenesis or via RNAi technique to determine the loss-of-function phenotype. A smaller subset will be pursued in great depth by examining interactions and cellular localization in vivo.

拟南芥中的多种信号传导网络利用异源三聚体G蛋白复合物将在细胞表面由G蛋白偶联受体（GPCR）感知的信号偶联到称为效应子的酶。到目前为止，下游效应物和上游GPCR都没有明确确定。这种信息的缺乏与人类和C.线虫G蛋白偶联途径，其中950个GPCR和十几个效应物已被确定，因此代表了后生动物中最广泛的信号网络。 我们将使用高通量的相互作用筛选来鉴定与异源三聚体G蛋白复合物相互作用的蛋白质。将测试相互作用物的体内相互作用，并将确认的候选物用作诱饵以启动额外的筛选轮。这些数据将进行生物信息学和图论荟萃分析，以确定相互作用网络中的主要信号转导途径和蛋白质复合物（信号体）。将更彻底地对选定的互动者进行调查。反向遗传学和分子生物学方法将被应用于确定这些元素在G蛋白信号网络的哪些分支上起作用。将通过插入诱变或通过RNAi技术敲除多达100个编码确认的相互作用物的基因，以确定功能丧失表型。通过检查体内相互作用和细胞定位，将深入研究较小的子集。