MOLECULAR MECHANISM OF MAMMALIAN AUTOPHAGY

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 自噬作为一种普遍存在的降解和回收蛋白质和细胞器所必需的溶酶体途径，已经涉及广泛的人类疾病，包括癌症、感染、肝病、肌病和神经变性。然而，组成哺乳动物自噬途径的分子机制以及这种机制如何发挥作用的分子机制在很大程度上是未知的。我们的长期目标是了解哺乳动物自噬途径的分子机制及其与人类疾病的相关性，以发现急需的早期诊断生物标志物和治疗靶点。为了实现这一目标，我们的总体目标是首先确定哺乳动物自噬途径中的新型蛋白质-蛋白质相互作用，然后确定这些新型相互作用因子的功能。Beclin 1是第一个被报道的哺乳动物自噬蛋白，在自噬调控中起着重要的积极作用，并与多种人类生理和病理有关，包括肿瘤抑制、神经退行性变、发育和衰老。因此，本研究将探讨Beclin 1介导的自噬调控，从而开始破译哺乳动物自噬的分子机制。在强有力的初步数据的指导下，三个具体的目标将采用一种新的综合小鼠遗传-蛋白质组学方法以及生物化学和细胞生物学研究的组合。目的一，我们将通过从活体动物中鉴定Beclin 1相互作用蛋白，建立一个环境依赖的哺乳动物自噬相互作用组的框架;目的二，我们将阐明两个新的Beclin 1相互作用蛋白Atg 14 L和Rubicon调控自噬的分子细节;在目标3中，我们将探索第三种新的Beclin 1相互作用物调控自噬的分子细节及其与癌症的相关性。通过建立哺乳动物自噬相互作用组的框架，并揭示Beclin 1及其相互作用蛋白调节自噬的分子细节，这项拟议的工作不仅将深入了解自噬在哺乳动物中的作用，而且还将为预防和治疗干预产生新的潜在目标，最终可能帮助患有自噬相关人类疾病的患者。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. As a ubiquitous lysosomal pathway essential for degrading and recycling proteins and organelles, autophagy has been implicated in a broad spectrum of human disease including cancer, infection, liver disease, myopathy and neurodegeneration. However, the molecular machinery comprising the mammalian autophagy pathway and the molecular mechanism of how this machinery functions are largely unknown. Our long-term goal is to understand the molecular mechanism of the mammalian autophagy pathway and its relevance to human disease, for discovering much needed early diagnostic biomarkers and therapeutic targets. To achieve this goal, our overall objective is to first identify novel protein-protein interactions in the mammalian autophagy pathway and subsequently to determine the functions of these novel interactors. Beclin 1, the first reported mammalian autophagy protein, plays an important positive role in autophagy regulation and has been implicated in a variety of human physiology and pathology, including tumor suppression, neurodegeneration, development and aging. Therefore, for this study, we will investigate Beclin 1-mediated autophagy regulation so as to begin deciphering the molecular mechanism of mammalian autophagy. Guided by strong preliminary data, three specific aims will be pursued using a combination of a novel integrated mouse genetic-proteomic approach as well as biochemical and cell biological studies. In Aim 1, we will establish the framework of a context-dependent mammalian autophagy interactome through identifying Beclin 1-interacting proteins from living animals; in Aim 2, we will elucidate the molecular details of autophagy regulation by two novel Beclin 1-interacting proteins, Atg14L and Rubicon; and in Aim 3, we will explore the molecular details of autophagy regulation by a third novel Beclin 1-interactor and its cancer relevance. By establishing the framework of the mammalian autophagy interactome and unraveling the molecular details of autophagy regulation by Beclin 1 and its interacting proteins, this proposed work will not only provide insight into the role of autophagy in mammals but also generate novel potential targets for preventive and therapeutic interventions that may ultimately aid patients suffering from autophagy-related human disease.