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Molecular Analysis of Tmie in sensory hair cells

感觉毛细胞中 Tmie 的分子分析

基本信息

批准号：
10395471
负责人：
Teresa A Nicolson
金额：
$ 33.26万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-05-04 至 2023-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10395471
关键词：
Address Affect Alleles Amino Acid Motifs Amino Acids Behavior Binding Biochemical Biology Calcium and Integrin Binding Protein 2 Cell physiology Cells Chimera organism Cochlea Collection Complex Data Dominant-Negative Mutation Ear Equilibrium Fluorescence Future Gene Proteins Gene Transfer Techniques Generations Genes Genetic Goals HMGA2 gene Hair Hair Cells Head Movements Hearing Human Image Integral Membrane Protein Knockout Mice Labyrinth Larva Length Light Lipoma Mechanics Mediating Membrane Membrane Proteins Methods Modeling Molecular Molecular Analysis Mouse Protein Mus Mutate Mutation Organ Outer Hair Cells PCDH15 gene Patients Pattern Peptide Signal Sequences Phenotype Play Protein Isoforms Proteins Role Seminal Sensory Sensory Hair Signal Transduction Site Stimulus Structure System Tertiary Protein Structure Testing Transgenes Transgenic Organisms Transmembrane Domain Two-Hybrid System Techniques Ubiquitin Validation Venus Vertebrates Vestibular Hair Cells Western Blotting Work Yeasts Zebrafish Zebrafish Proteins base cell type cellular transduction deafness experimental study extracellular gene function in vivo insight lateral line link protein mechanotransduction member mutant novel protein complex protein protein interaction sensor sound tool

项目摘要

PROJECT SUMMARY Transmembrane inner ear-expressed gene (TMIE) encodes a transmembrane protein that essential for hearing and vestibular function in vertebrates. A recent study of Tmie-/- mice has provided strong evidence for a central role of TMIE in mechanotransduction in cochlear hair cells. As a member of the transduction complex, mouse TMIE can bind to a specific isoform of Protocaderin 15 and/or another membrane protein, protein Lipoma HMGIC Fusion Partner-Like 5 (LHFPL5). Despite these seminal findings, it is not well understood how TMIE functions as a member of the transduction complex, and to date, its role in vestibular hair cells is unexplored. The aim of the present proposal is to conduct a comprehensive study of zebrafish Tmie in terms of identifying the protein motifs that are required for (i) localization of Tmie to stereocilia, (ii) modulation of mechanotransduction, and (iii) genetic and biochemical interactions with other members of the transduction complex. The proposed experiments will take advantage of our collection of mechanotransduction mutants and tools we have developed in zebrafish to investigate the function of Tmie in hair cells. Our preliminary data indicate that zebrafish Tmie is required the localization of Transmembrane channel like 1 (Tmc1) and Tmc2b to the stereocilia of hair cells in the inner ear and lateral line organ. This observation is surprising in light of experiments with Myc tagged TMC2 in mouse outer hair cells, however, the results suggest that TMIE may play different roles in different cell types. The proposed experiments will expand upon these novel findings and identify the amino acid motifs in Tmie that promote interaction with Lhfpl5a and the localization of the Tmcs to the site of mechanotransduction in hair cells. This work will provide mechanistic insights into the role of Tmie in hair cells and provide a better understanding the molecular determinants of Tmie that are critical for vertebrate hearing and balance.

项目概要跨膜内耳表达基因 (TMIE) 编码对听力至关重要的跨膜蛋白和脊椎动物的前庭功能。最近对 Tmie-/- 小鼠的一项研究为 TMIE 在耳蜗毛细胞机械转导中的核心作用。作为转导复合体的成员，小鼠 TMIE 可以与 Protocaderin 15 的特定亚型和/或另一种膜蛋白、蛋白质结合脂肪瘤 HMGIC 融合伙伴样 5 (LHFPL5)。尽管有这些开创性的发现，但尚不清楚如何 TMIE 作为转导复合体的成员发挥作用，迄今为止，它在前庭毛细胞中的作用是未经探索。本提案的目的是对斑马鱼 Tmie 进行全面研究鉴定 (i) Tmie 定位到静纤毛，(ii) 调节所需的蛋白质基序机械转导，以及（iii）与转导其他成员的遗传和生化相互作用复杂的。拟议的实验将利用我们收集的机械转导突变体和我们在斑马鱼中开发了工具来研究 Tmie 在毛细胞中的功能。我们的初步数据表明斑马鱼 Tmie 需要像 1 (Tmc1) 和 Tmc2b 这样的跨膜通道的定位内耳和侧线器官中毛细胞的静纤毛。这一观察结果令人惊讶在小鼠外毛细胞中进行 Myc 标记的 TMC2 实验，然而，结果表明 TMIE 可能在不同的细胞类型中发挥不同的作用。拟议的实验将扩展这些新的发现和鉴定 Tmie 中促进与 Lhfpl5a 相互作用的氨基酸基序以及 Tmc 的定位毛细胞中机械传导的部位。这项工作将为 Tmie 在毛细胞并提供更好的理解 Tmie 的分子决定因素，这对脊椎动物至关重要听力和平衡能力。