Mechanisms of Protein Assembly and Transport in Hair Cells

毛细胞中蛋白质组装和运输的机制

基本信息

批准号：
10290317
负责人：
Christopher L Cunningham
金额：
$ 15.75万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-12-01 至 2023-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10290317
关键词：
Actins Affect Apical Binding Binding Proteins Biochemical Brain CRISPR/Cas technology Ca(2+)-Transporting ATPase Catechol O-Methyltransferase Cell surface Cells Co-Immunoprecipitations Cochlea Communication Complement Complementary DNA Data Defect Disease Endoplasmic Reticulum Endosomes Epithelial Cells Epitopes Fluorescence Microscopy Foundations Gene Targeting Genes Goals Golgi Apparatus Hair Cells Health Homologous Gene Human Impairment Individual Integral Membrane Protein Ion Channel Ion Channel Gating Label Length Libraries Link Mechanics Mediating Methyltransferase Molecular Morphology Mus Mutant Strains Mice Mutation Mutation Analysis Organelles Pathway interactions Point Mutation Process Property Proteins Quality of life Resolution Role Route Sensory Set protein Signal Transduction Structure Surface Synapses Tertiary Protein Structure Testing Tissues Transmission Electron Microscopy Validation Yeasts Zebrafish auditory processing base deafness experimental study hearing impairment insight loss of function mutation mechanical stimulus mechanotransduction molecular marker mutant polarized cell protein protein interaction protein transport sound therapeutic development tool trafficking yeast two hybrid system

项目摘要

Project Summary/Abstract The overall goal of the project is to define and characterize the organelles and specialized molecular machinery for protein assembly, processing and transport in hair cells, and the defects in this process that cause deafness. We propose here to investigate the mechanisms by which TOMT regulates transport of a subset of functionally important proteins in hair cells, and to begin to uncover TOMT-associated proteins that participate. Based on our preliminary data, we hypothesize that TOMT is an endoplasmic reticulum protein that is critical for assembly and transport of a specific set of transmembrane proteins in hair cells. To test our hypothesis, we will: i) use immunolocalization and immunogold transmission electron microscopy to precisely localize TOMT to hair-cell specific transport organelles; ii) identify and characterize important TOMT functional domains; and iii) look for interaction partners for TOMT that functionally cooperate in hair cell protein transport. Our preliminary data show the feasibility of our approach. We anticipate that our studies will elucidate new insights into the unique molecular machinery of hair cell protein transport that establishes distinctive functional specializations critical for auditory processing. Furthermore, a better understanding of protein transport mechanisms in hair cells has the potential to provide important context for protein transport deficits that are common in hearing loss.

项目摘要/摘要该项目的总体目标是定义和表征细胞器和专业分子蛋白质组装，加工和运输毛细胞的机械以及此过程中的缺陷引起耳聋。我们在这里建议调查TOMT调节A运输的机制毛细胞中功能上重要蛋白的子集，并开始发现与TOMT相关的蛋白参加。根据我们的初步数据，我们假设TOMT是一种内质网蛋白对于毛细胞中一组特定的跨膜蛋白的组装和运输至关重要。测试我们的假设，我们将：i）使用免疫定位和免疫金传输电子显微镜精确将Tomt定位到发型特定的运输细胞器中； ii）识别并表征重要的TOMT功能域； iii）寻找在毛细胞蛋白运输中功能合作的TOMT的相互作用伙伴。我们的初步数据显示了我们方法的可行性。我们预计我们的研究将阐明新的深入了解毛细胞蛋白转运的独特分子机制，该机械建立了独特的功能对听觉处理至关重要的专业。此外，对蛋白质转运的更好理解毛细胞中的机制有可能为蛋白质转运缺陷提供重要背景听力损失常见。