Quality control of the primary cilium proteome

初级纤毛蛋白质组的质量控制

• 批准号: 10546935
• 负责人: Maxence V Nachury
• 金额: $ 5.35万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10546935
• 关键词: Appearance; Bardet-Biedl Syndrome; Binding; Biological Assay; Cell membrane; Cells; Cilia; Complex; Coupled; Cues; Cystic kidney; Data; Development; Diffuse; Diffusion; Disease; Endocytosis; Endosomes; Erinaceidae; Etiology; Event; Excision; Functional disorder; Funding; G-Protein-Coupled Receptors; Goals; Hair; Health; Hereditary Disease; Homeostasis; Human; Investigation; Knowledge; Lateral; Lead; Ligase; Light; Link; Membrane Proteins; Modeling; Molecular; Movement; Myosin ATPase; Obesity; Pathway interactions; Polydactyly; Proteins; Proteome; Proteomics; Quality Control; Reader; Receptor Signaling; Regulation; Research; Rest; Retinal Degeneration; Role; SSTR3 gene; Signal Pathway; Signal Transduction; Smell Perception; Sorting - Cell Movement; Structure; Testing; Therapeutic Intervention; Ubiquitin; Ubiquitination; Variant; Vision; Work; base; hedgehog signal transduction; kidney malformation; molecular imaging; mutant; novel; programs; protein complex; recruit; sensor; sensory input; single molecule; skeletal abnormality; smoothened signaling pathway; trafficking; ubiquitin ligase

PROJECT SUMMARY Primary cilia organize signaling pathways such as vision, olfaction and Hedgehog signaling. The movements of signaling receptors into, inside and out of cilium are critical for the correct regulation of these pathways, yet our understanding of the basic mechanisms governing signaling receptor trafficking through cilia remains fragmentary. Past work from the lab identified and characterized the BBSome, a protein complex that ferries signaling receptors out of cilia. The relevance of the BBSome to human health and disease is evidence by the fact that BBSome dysfunction causes Bardet-Biedl Syndrome (BBS), a hereditary disease characterized by obesity, retinal degeneration, polydactyly and kidney malformations. The major goal of this proposal is to determine how the BBSome selects signaling receptors for removal from cilia. The emphasis in this funding period will be on investigating the role of ubiquitin in tagging membrane proteins for removal from cilia. Preliminary data indicate the existence of a ciliary machinery that recognizes activated GPCRs, ubiquitinates them and sorts ubiquitinated GPCRs out of cilia. We will characterize the molecules acting at each of these steps using quantitative assays for signal-dependent GPCR exit. The hierarchical ordering of the molecular cogs and levers that affix and read ubiquitin on proteins that need to exit cilia promises to uncover a multi-step pathway reminiscent of the ESCRT machinery responsible for degradative sorting. Finally, the fate of GPCRs that exit cilia will be tracked by single-molecule imaging to determine whether endocytosis is coupled to ciliary exit or whether GPCRs instead diffuse into the plasma membrane after exiting cilia. The proposed studies will cast new light on how the ciliary abundance of proteins is regulated and open the door to a mechanistic investigation of ciliary quality control.

项目摘要 原发性纤毛组织信号通路，例如视觉，嗅觉和刺猬信号传导。这 信号受体进入，内外的纤毛运动对于正确调节至关重要 途径，但是我们对管理信号受体运输的基本机制的理解 仍然是碎片的。实验室的过去工作确定并表征了BBSOME，这是一种蛋白质复合物 渡轮信号受体来自纤毛。 BBSOME与人类健康和疾病的相关性是证据 BBSOME功能障碍会导致Bardet-Biedl综合征（BBS），这是一种遗传性疾病 通过肥胖，视网膜变性，多态和肾脏畸形。 该提案的主要目标是确定BBSOME如何选择信号受体以拆卸 来自Cilia。在此资金期间的重点将是研究泛素在标记膜上的作用 从纤毛中去除的蛋白质。初步数据表示存在识别的睫状机械的存在 激活的GPCR，将它们泛素化，并从Cilia中分类泛素化的GPCR。我们将表征 使用定量测定对信号依赖性GPCR出口作用的分子。这 分子齿轮和杠杆的分层排序，这些齿轮和杠杆粘合并读取需要退出的蛋白质上的泛素 西里亚（Cilia）承诺要揭露多步途径 降解分类。最后，退出CILIA的GPCR的命运将通过单分子成像跟踪 确定内吞作用是否与睫状出口耦合，还是GPCR散布到等离子体中 退出纤毛后的膜。 拟议的研究将为如何调节蛋白质的睫状丰度并打开新的启示。 对睫状质量控制的机械调查的门。