Spatial Regulation of RGS and G Protein Signaling

RGS 和 G 蛋白信号传导的空间调控

基本信息

批准号：
7389129
负责人：
MARILYN Gist FARQUHAR
金额：
$ 5.51万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-04-17 至 2009-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7389129
关键词：
Abbreviations Adrenergic Receptor Binding Biochemical Bioinformatics Biological Assay Biological Process Cell physiology Clathrin Clathrin-Coated Vesicles Coated vesicle Collaborations Complex Data Development Dissociation EGF gene Early Endosome Electron Microscopy Endocytic Vesicle Endoplasmic Reticulum Epidermal Growth Factor Receptor Event Fibrinogen Fingers G Protein-Coupled Receptor Genes G-Protein Signaling Pathway GTP-Binding Protein Regulators GTP-Binding Proteins GTPase-Activating Proteins Goals Growth Growth Factor Growth Factor Receptors Guanine Guanine Nucleotide Dissociation Inhibitors Human Immunoelectron Microscopy Immunofluorescence Immunologic Insulin-Like Growth Factor I Knowledge Link Localized Location Mediating Microscopy Mitogen-Activated Protein Kinases Modeling Molecular NGFR Protein Nerve Growth Factor 1 Nerve Growth Factor Pathway Nerve Growth Factors Numbers Opioid Receptor PC12 Cells PDZ protein Pathway interactions Phosphatidylinositols Phospholipid Metabolism Phylogenetic Analysis Process Program Research Project Grants Protein Binding Protein C Protein Overexpression Proteins Proteomics RGS Domain RGS Proteins RGS19 gene RNA Interference Receptor Down-Regulation Receptor Signaling Regulation Role Sequence Analysis Signal Pathway Signal Transduction Stimulation of Cell Proliferation Tertiary Protein Structure Testing Time Work antitumor agent base cell growth coated pit comparative fly human RGS19 protein inhibitor/antagonist knock-down multicatalytic endopeptidase complex mutant novel protein degradation receptor research study stem trafficking ubiquitin-protein ligase uptake yeast two hybrid system

项目摘要

During the last five years, we identified several new proteins that are components of novel G protein signaling pathways. Among them are RGS proteins that not only serve as GAPs for G proteins through their RGS domains, but also bind to other proteins through their unique N- and C-termini. We have concentrated on two RGS proteins, GAIP (RGS19), a GAP for Go_i3, and RGS-PX1, a GAP for Go_s which also has a PX domain that identifies it as it as a SNX protein involved in phosphoinositide signaling. We have obtained results indicating that RGS proteins link G protein signaling to other signaling pathways, including EGF and TrkA growth factor signaling and phosphoinositide signaling, and to other cell processes such as protein degradation. The overall goal of the proposed work is to define the molecular components of the novel signalin! networks we have uncovered and to establish their cellular location, trafficking and functions. The underlyin themes are that RGS proteins are both GAPs and bddging molecules that link signaling events and that these interactions are spatially regulated by ceHuJar distribution and trafficking. We will focus on three specific aims: Specific Aim #1: To assess the role of RGS-PX1 in tinking Go_s signaling and EGF receptor (EGFR) down-regulation at early endosomes. This will be accomplished by assessing the effects of activating the _2- adrenergic receptor and G(zs on EGFR degradation, MAP kinase activation, RGS-PX1 localization and, distribution of Go_s-GFP. We hypothesize that RGS-PX1 serves to link signals generated through activation of I Go_sto EGFR signaling. Specific Aim #2: To determine if GIPC functions to organize signaling complexes that I function in growth factor signaling and trafficking. GIPC, a PDZ protein, binds both GAIP and the TrkA growth I factor receptor. We wUi use deconvolution an(] immunoelectron microscopy, a proteomics approach and RNAi I experiments to investigate GIPC s role in assembiy of signaling and trafficking complexes. Specific Aim #3: I To further define Gczi3-GAIP-GIPC-GIPN signaling networks by biochemical, morphological and bioinformatics I approaches= We will concentrate here on i) isolation of putative signaling complexes containing GAIP, G(zi3,j and GPCR, ii) characterization of GIPN, a novel GAIP-interacting protein that is a putative E3 ligase, and iii) usel of bioinformatics to model God-GAIP-GIPC/GIPN signaling networks. These studies can be expected to I considerably expand knowledge of the role of RGS proteins in linking G protein and growth factor signaling. ! Due to their broad functions in regulation of cell functions, including cell growth, differentiation and mitogenesis, t RGS proteins represent attractive targets for development of pharmacologic and anti-tumor agents. 1

在过去的五年中，我们确定了几种新蛋白，它们是新型G蛋白的成分信号通路。其中包括RGS蛋白，不仅可以充当G蛋白的差距 RGS结构域，但也通过其独特的N-和C末端结合到其他蛋白质。我们专注于两个RGS蛋白GAIP（RGS19），GO_I3的差距和RGS-PX1，GO_S的差距也有PX 将其识别为参与磷酸肌醇信号传导的SNX蛋白的结构域。我们已经获得了结果表明RGS蛋白将G蛋白信号传导与其他信号通路联系起来，包括EGF和TRKA 生长因子信号传导和磷酸肌醇信号传导，以及其他细胞过程，例如蛋白质降解。拟议的工作的总体目标是定义新颖信号的分子成分！我们发现了网络，并建立了其细胞位置，贩运和功能。基础主题是RGS蛋白既是连接信号事件的间隙和bddging分子，又是这些分子相互作用在空间上通过Cehujar的分布和贩运来调节。我们将专注于三个具体目标：特定目的＃1：评估RGS-PX1在Tinking GO_S信号传导和EGF受体（EGFR）中的作用早期内体的下调。这将通过评估激活_2-的影响来实现。肾上腺素能受体和G（Zs on EGFR降解，MAP激酶激活，RGS-PX1定位以及， GO_S-GFP的分布。我们假设RGS-PX1用于通过激活I的链接信号 GO_STO EGFR信令。特定目的＃2：确定GIPC是否功能来组织信号复合体I 生长因子信号传导和运输中的功能。 GIPC是PDZ蛋白，结合GAIP和TRKA生长I 因子受体。我们使用反卷积AN（]免疫电子显微镜，一种蛋白质组学和RNAi I 研究GIPC在信号传导和运输复合物组装中的作用的实验。特定目标＃3：我进一步定义了通过生化，形态学和生物信息学的GCZI3-GAIP-GIPC GIPC信号网络方法=我们将集中精力i）分离含有GAIP的推定信号传导复合物（Zi3，j）和GPCR，ii）GIPN的表征，GIPN是一种新型的Gaip相互作用蛋白，是一种推定的E3连接酶，iii）Usel 生物信息学的建模，以建模上帝GAIP GIPC/GIPN信号网络。这些研究可以预期大大扩展了RGS蛋白在连接G蛋白和生长因子信号传导中的作用的知识。呢由于它们在调节细胞功能方面的广泛功能，包括细胞生长，分化和有丝分裂发生，t RGS蛋白代表了用于开发药理学和抗肿瘤剂的有吸引力的靶标。 1