Global analysis of bundle protein transport in mouse vestibular hair cells

描述（由申请人提供）：声音和平衡的感知取决于毛束的机械感觉功能，毛束是一组从听觉和前庭毛细胞顶面伸出的充满肌动蛋白的静纤毛。对与毛发相关的基因的研究已经鉴定出许多定位于毛束并有助于其发育和功能的蛋白质[1]。尽管取得了这些进展，但构成毛束并调节其成熟、维持和功能的完整蛋白质网络仍然定义不清。虽然蛋白质特异性运输到束的机制也知之甚少，但积累的证据表明肌球蛋白马达蛋白可能发挥重要作用[2]。这项提议的目的是鉴定束蛋白 在毛束形成过程中，这些蛋白质被运输并维持在毛束内，并检验肌球蛋白-VIIa（MYO 7A）在这些蛋白质的运输中起作用的假设。使用蛋白质组学方法以及其他生物化学技术，本提案中的目标将1）以无偏的方式鉴定在毛束组装期间和之后在毛束内富集的蛋白质网络，2）确定MYO 7A是否在特定蛋白质向毛束的运输中起作用，以及3）鉴定这些毛束特异性蛋白质中的哪一种直接或间接地与MYO 7A结合。 与MYO 7A相互作用。这些研究的结果将大大有助于我们对毛束组装和功能的分子理解，并有助于指导毛细胞修复和再生治疗策略的发展。

DESCRIPTION (provided by applicant): The perception of sound and balance depends on the mechanosensory function of the hair bundle, a group of actin-filled stereocilia that project from the apical surface of auditory and vestibular hair cells. The study of deafness-linked genes has led to the identification of a number of proteins that are localized to the hair bundle and contribute to its development and function [1]. Despite these advances, the complete network of proteins that constitute the hair bundle and regulate its maturation, maintenance and function remain poorly defined. Although the mechanisms by which proteins are specifically transported to the bundle are also poorly understood, accumulated evidence suggests that myosin motor proteins may play important roles [2]. The goal of this proposal is to identify the bundle proteins that are transported to and maintained within the hair bundle during its formation and to test the hypothesis that myosin-VIIa (MYO7A) plays a role in the transport of these proteins. Using proteomics approaches as well as other biochemical techniques, the aims in this proposal will 1) identify in an unbiased manner the protein networks that are enriched within the hair bundle during and after bundle assembly, 2) determine whether MYO7A plays a role in the transport of specific proteins to the bundle and 3) identify which of these bundle-specific proteins directly or indirectly interact with MYO7A. The results of these studies will significantly contribute to our molecular understanding of bundle assembly and function and could help guide the development of therapeutic strategies for hair cell repair and regeneration.