MOLECULAR MECHANISM OF MAMMALIAN AUTOPHAGY

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 作为一种无处不在的溶酶体途径，自噬对蛋白质和细胞器的降解和循环是必不可少的，它与多种人类疾病有关，包括癌症、感染、肝病、肌病和神经退行性变。然而，组成哺乳动物自噬途径的分子机制以及该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新的蛋白质-蛋白质相互作用，然后确定这些新的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，在自噬调节中发挥着重要的积极作用，并参与了多种人类生理和病理过程，包括肿瘤抑制、神经退行性变、发育和衰老。因此，在本研究中，我们将研究Beclin 1介导的自噬调控，以期开始破译哺乳动物自噬的分子机制。在强大的初步数据的指导下，将结合新的小鼠遗传-蛋白质组学方法以及生化和细胞生物学研究来追求三个具体目标。在目标1中，我们将通过从活体动物中鉴定Beclin 1相互作用蛋白来建立与上下文相关的哺乳动物自噬相互作用体的框架；在目标2中，我们将阐明两个新的Beclin 1相互作用蛋白Atg14L和Rubcon调控自噬的分子细节；在目标3中，我们将探索第三个新的Beclin 1相互作用蛋白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体的框架，并解开Beclin 1及其相互作用蛋白调控自噬的分子细节，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，还将为预防和治疗干预创造新的潜在靶点，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. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. 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.