Mechanisms of Protein Assembly and Transport in Hair Cells

项目摘要/摘要 该项目的总体目标是定义和描述细胞器和专门的分子 毛细胞中蛋白质组装、加工和运输的机制，以及这一过程中的缺陷 导致耳聋。我们建议在这里研究TOMT调节细胞转运的机制。 毛细胞中具有重要功能的蛋白质的子集，并开始发现与TOMT相关的蛋白质 参与进来。根据我们的初步数据，我们假设TOMT是一种内质网蛋白， 对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透射电子显微镜来精确 将TOMT定位于毛细胞特异性运输细胞器；ii)鉴定和表征重要的TOMT功能 以及iii)为TOMT寻找在毛细胞蛋白质运输方面进行功能合作的相互作用伙伴。 我们的初步数据表明了我们方法的可行性。我们预计，我们的研究将阐明新的 对毛细胞蛋白质运输的独特分子机制的洞察建立独特的功能 对听觉处理至关重要的专门化。此外，更好地了解蛋白质的运输 毛细胞中的机制有可能为蛋白质运输缺陷提供重要的背景 常见于听力损失。

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.