Structure and Function of the Ciliary Pore Complex

睫状孔复合体的结构和功能

• 批准号: 8977649
• 负责人: Kristen J. Verhey
• 金额: $ 50.31万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-15 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8977649
• 关键词: Abbreviations; Adult; Automobile Driving; Binding; Biological; Carrier Proteins; Cell physiology; Cell surface; Cells; Cilia; Cilium Microtubule; Complement; Complex; Cystic Kidney Diseases; Data; Defect; Development; Dimensions; Disease; Electron Microscopy; Erinaceidae; Event; Flagella; Fluorescence Recovery After Photobleaching; Growth Factor; Image; Importins; Joubert syndrome; Kinesin; Lateral; Lead; Left; Life; Ligands; Link; Lipids; Lung; Membrane; Membrane Proteins; Methods; Microscopy; Microtubules; Modeling; Molecular; Monomeric GTP-Binding Proteins; Motor; Movement; Nature; Nephronophthisis; Nuclear; Nuclear Pore; Nuclear Pore Complex; Nuclear Pore Complex Proteins; Organ; Organelles; Phenotype; Physiology; Plant Roots; Play; Precipitation; Process; Proteins; Publishing; Resolution; Retinitis Pigmentosa; Role; Running; Sensory; Signal Transduction; Structure; Syndrome; Techniques; Testing; Therapeutic; Work; base; body system; cell motility; ciliopathy; human SMO protein; human disease; novel; particle; protein protein interaction; protein transport; public health relevance; single molecule; spatial relationship; spatiotemporal; tool; trafficking; yeast two hybrid system

 DESCRIPTION (provided by applicant): Cilia are microtubule-based organelles that are involved in multiple cellular processes including signaling and motility. Protein entry into cilia s a tightly regulated process and defects in cilia-localized proteins are felt to be a root cause of many ciliopathies that manifest as cystic kidney disease, retinitis pigmentosa and/or other defects. The entry of proteins into cilia appears to be controlled at the cilia base where a ciliar pore is hypothesized to exist. Our group has proposed that the ciliary pore is highly analogous to the nuclear pore in its molecular composition and regulatory mechanism. In particular, we have demonstrated that proteins known to be involved in nuclear trafficking - the small GTPase Ran, its binding partners the importins, and nucleoporins - are present at the base or within cilia and regulate protein entry into the ciliary compartment. Based on our Preliminary Data, we now propose a model where two distinct mechanisms regulate entry into the ciliary compartment: entry of cytosolic proteins is regulated by nucleoporins of the ciliary pore complex (CPC) and requires kinesin motors and intraflagellar transport (IFT) whereas entry of membrane proteins is regulated by molecules of the transition zone (TZ) and is IFT-independent. We will test this model using inducible inhibition of nucleoporin and kinesin motor function combined with live-cell, single-molecule, and super-resolution microscopy techniques. These studies will lead the field forward in understanding ciliary gating mechanisms and will make important inroads in defining the nature of the cilia pore and its relation to the nuclear pore.

 描述(申请人提供)：纤毛是以微管为基础的细胞器，参与多种细胞过程，包括信号和运动。纤毛蛋白进入纤毛S纤毛定位蛋白的严格调控过程和缺陷被认为是许多纤毛疾病的根本原因，这些疾病表现为囊性肾病、视网膜色素变性和/或其他缺陷。蛋白质进入纤毛似乎受到纤毛底部的控制，纤毛底部假设存在纤毛孔。我们的研究小组提出，睫状孔在其分子组成和调节机制上与核孔高度相似。特别是，我们已经证明了已知的与核运输有关的蛋白质-小gtp酶ran、其结合伙伴重要蛋白和核孔蛋白-存在于碱基或纤毛内。 并调节进入睫状室的蛋白质。根据我们的初步数据，我们现在提出了一个模型，其中有两种不同的机制调节进入纤毛室：胞浆蛋白的进入由睫状孔复合体的核孔蛋白(CPC)调节，需要动蛋白马达和鞭毛内转运(IFT)，而膜蛋白的进入由过渡区(TZ)的分子调节，不依赖于IFT。我们将结合活细胞、单分子和超分辨率显微镜技术，使用对核孔素和运动蛋白运动功能的诱导抑制来测试这一模型。这些研究将引领该领域在理解纤毛门控机制方面取得进展，并将在确定纤毛孔的性质及其与核孔的关系方面取得重要进展。