Mechanisms of Ciliary Trafficking of the Olfactory CNG Channel

嗅觉 CNG 通道的纤毛运输机制

• 批准号: 7545606
• 负责人: Paul Michael Jenkins
• 金额: $ 3.19万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7545606
• 关键词: 8-bromocyclic GMP; Abbreviations; Acidic Amino Acids; Action Potentials; Address; Affect; Afferent Neurons; Alanine; Anosmia; Binding; Biological Assay; Biological Models; Brain; Caenorhabditis elegans; Cations; Cell physiology; Cells; Chemicals; Cilia; Complex; Cyclic AMP; Cyclic GMP; Cyclic Nucleotides; Defect; Disease; Distal; Dominant-Negative Mutation; Dynein ATPase; Epithelial Cells; Etiology; Family member; Flagella; Functional disorder; GTP-Binding Proteins; Goals; Homologous Gene; Human; Image; Infertility; Invertebrates; Ion Channel; KRP protein; Kinesin; Knockout Mice; Knowledge; Laboratories; Lead; MDCK cell; Martens; Mediating; Microtubules; Motor; Movement; Mus; Nose; Numbers; Odorant Receptors; Olfactory Epithelium; Organelles; PKD2 protein; Pathway interactions; Phosphorylation; Play; Polycystic Kidney Diseases; Process; Protein Binding; Protein Sortings; Proteins; Publishing; Retinitis Pigmentosa; Role; Scanning; Sensory; Serine; Signal Transduction; Signaling Molecule; Signaling Protein; Sorting - Cell Movement; Structure; System; Testing; Threonine; Tissues; Translations; Viral; Viral Physiology; Work; adenylyl cyclase III; anterograde transport; base; casein kinase II; cyclic-nucleotide gated ion channels; design; human disease; inhibitor/antagonist; insight; mutant; protein B; protein transport; research study; respiratory; retrograde transport; small hairpin RNA; trafficking

DESCRIPTION (provided by applicant): The overall goal of this proposal is to develop a better understanding of the mechanisms underlying protein transport into olfactory sensory neuron (OSN) cilia. Cilia are complex, microtubule-rich, organelles uniquely adapted for diverse cellular functions. Defects in cilia structure and function have been shown to underlie a number of human diseases. Sensory cilia on dendritic endings of OSNs compartmentalize signaling molecules, including cyclic nucleotide-gated (CNG) channels that are the primary targets of odorant-induced signaling. Olfactory CNG channels are responsible for the translation of the chemical signal from the odorant receptor into action potentials that are processed in the brain. Despite a wealth of knowledge about CNG channel structure and function, little is known about the mechanisms of their subcellular targeting. Movement of proteins within the cilia is governed by intraflagellar transport (IFT), an evolutionary conserved process that facilitates movement of cargo along cilia and flagella. In mammalian systems, the heterotrimeric Kinesin II plays a clear role in IFT; however, a role in ciliary transport has not yet been established for the homodirrieric KIF17. Newly published experiments I performed in Dr. Martens' laboratory have shown that KIF17 is required for ciliary enrichment of the olfactory CNG channel in a heterologous system; however, the role for KIF17 in native olfactory epithelium remains unknown. Recently it has been shown that the proper targeting of another ciliary protein, nephrocystin 1, in nasal respiratory epithelial cells requires the binding of phosphofurin acidic cluster sorting protein 1 (PACS-1). PACS-1 is an intracellular sorting protein that binds to phosphorylated serine residues contained in a cluster of acidic residues on the cargo protein. PACS-1 has been shown to interact with several other acidic cluster-containing ion channel, however no role has been defined for PACS-1 in olfactory cilia transport. It is my hypothesis that the ciliary targeting of olfactory CNG channels is dependent on both KIF17 and PACS-1 in native olfactory epithelium. The following specific aims are designed to test this hypothesis: Specific Aim 1: To investigate the role of KIF17 in the ciliary transport of the olfactory CNG channel in native olfactory sensory neurons. Specific Aim 2: To determine the effects of PACS-1 on CNG channel ciliary targeting. I will address these aims using confocal imaging, assays of olfactory function, and viral expression systems. My proposal provides a plan to further elucidate the mechanisms of ciliary enrichment of the olfactory CNG channel. Characterizing the general mechanisms of channel targeting will provide a better understanding of ciliary transport and give us further insight into the underlying etiologies of human ciliary disorders.

描述（由申请人提供）：本提案的总体目标是更好地了解蛋白质转运到嗅觉感觉神经元（OSN）纤毛的潜在机制。纤毛是一种复杂的、富含微管的细胞器，具有独特的细胞功能。纤毛结构和功能的缺陷已被证明是许多人类疾病的基础。OSN树突末端的感觉纤毛将信号分子区室化，包括作为气味诱导信号传导的主要靶点的环核苷酸门控（CNG）通道。嗅觉CNG通道负责将来自气味受体的化学信号翻译成在大脑中处理的动作电位。尽管对CNG通道的结构和功能有丰富的了解，但对其亚细胞靶向机制知之甚少。蛋白质在纤毛内的运动受鞭毛内转运（IFT）的控制，这是一个进化上保守的过程，有利于货物沿着纤毛和鞭毛的运动。在哺乳动物系统中，异源三聚体驱动蛋白II在IFT中起着明确的作用;然而，同源二聚体KIF 17在纤毛转运中的作用尚未确定。我在Martens博士实验室进行的新发表的实验表明，KIF 17是异源系统中嗅觉CNG通道纤毛富集所必需的;然而，KIF 17在天然嗅觉上皮中的作用仍然未知。最近，它已被证明，鼻呼吸道上皮细胞中的另一种纤毛蛋白，肾囊蛋白1的适当靶向需要磷酸弗林酸性簇分选蛋白1（PACS-1）的结合。PACS-1是一种细胞内分选蛋白，其与货物蛋白上的酸性残基簇中所含的磷酸化丝氨酸残基结合。PACS-1已被证明与其他几种含酸性簇的离子通道相互作用，但PACS-1在嗅觉纤毛转运中的作用尚未确定。这是我的假设，纤毛的嗅觉CNG通道的目标是依赖于天然嗅上皮中的KIF 17和PACS-1。具体目的1：研究KIF 17在天然嗅觉感觉神经元中嗅觉CNG通道纤毛转运中的作用。具体目的2：确定PACS-1对CNG通道纤毛靶向的影响。我将使用共聚焦成像，嗅觉功能测定和病毒表达系统来解决这些问题。我的建议提供了一个计划，以进一步阐明纤毛富集的嗅觉CNG通道的机制。表征通道靶向的一般机制将提供对纤毛运输的更好理解，并使我们进一步了解人类纤毛疾病的潜在病因。