Ubiquitin-dependent mitochondrial quality control

泛素依赖性线粒体质量控制

基本信息

批准号：
10369587
负责人：
MARIUSZ KARBOWSKI
金额：
$ 33.67万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-01 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10369587
关键词：
ATP phosphohydrolase Address Affect Alzheimer&apos s Disease Amyotrophic Lateral Sclerosis Autophagocytosis Biochemical Bioenergetics Biological Assay Biology Cell model Cells Complex Complications of Diabetes Mellitus Data Dependence Development Disease Equilibrium Eukaryotic Cell Genes Goals HCT116 Cells Hela Cells Homeostasis Huntington Disease Image In Vitro Laboratories Lead Link Mediating Membrane Proteins Messenger RNA Methods Mitochondria Mitochondrial Membrane Protein Mitochondrial Proteins Molecular Nerve Degeneration Neurons Outer Mitochondrial Membrane Parkin Parkinson Disease Pathology Pathway interactions Process Proteins Proteomics Publishing Quality Control Reactive Oxygen Species Regulation Role Signal Transduction Site Stress Structure System Tertiary Protein Structure Testing Time Ubiquitin Ubiquitination Work age related aging population biological adaptation to stress human disease improved insight mitochondrial dysfunction multicatalytic endopeptidase complex new therapeutic target novel prevent prospective protein degradation proteostasis success therapy development trafficking trend ubiquitin-protein ligase

项目摘要

Summary Mitochondria are at the center of age-related human diseases, such as Parkinson’s, Alzheimer’s, Huntington’s diseases. With current trends toward the aging population, developing therapies for these diseases is a critical need. Protective strategies, mostly aimed at neutralizing toxic reactive oxygen species (ROS), have only been partially successful. Thus, new treatment options with independent modes of action are needed to improve the success rate of current therapies. The proposed work directly responds to this need and will provide critical insights into the regulation and function of mitochondrial quality control pathways. Eukaryotic cells have a multi-layered system dedicated to mitochondrial quality control. The initial defense consists of a network of proteolytic systems that degrade proteins that are damaged, misfolded or mislocalized, or control activities of stress responsive mitochondrial factors. Recent findings, including work in the PI’s laboratory, demonstrated that the ubiquitin (Ub)/proteasome system (UPS), through Ub-dependent degradation/control of the OMM associated proteins is critical for mitochondrial quality control. Regulation of mitochondrial fusion and fission (mitochondrial dynamics) is another principal mitochondrial quality control mechanism. Recently we proposed that stability of outer mitochondrial membrane (OMM) proteins, including mitochondrial fusion factors Mfn1 and Mfn2 is controlled by mitochondrial fission proteins Drp1 and Mff. Consistent with this, our new data indicate that previously unsuspected Ub-dependent signaling functions of Drp1 and Mff are required for this process. We also found that mitochondrial protein ubiquitination is focal and that this process is accelerated in Mff-/- and Drp1-/- cells. These and other preliminary results, suggest unappreciated mechanisms regulating and coordinating mitochondrial quality control pathways, including participation of the novel mitochondrial protein degradation intermediates and mutual dependence between mitochondrial dynamics and mitophagy. We also identified an OMM-associated E3 Ub ligase Rnf179, which according to our preliminary findings not only controls stability of OMM proteins, including Mfn1 and Mfn2, but also affects mitochondria-specific autophagy (mitophagy), a critical UPS-dependent mitochondrial quality control pathway. The principle goal of this proposal is to test the hypothesis that through Ub-dependent signaling, Drp1 and Mff control and coordinate distinct mitochondrial quality pathways, including mitochondrial fission and fusion rates and activation of mitophagy. We also anticipate defining the role and mechanism of Rnf179 with mitochondrial protein ubiquitination in the control of mitochondrial function and integration of mitochondrial quality control pathways, including Parkin- dependent and -independent mitophagy. To achieve these goals we combine biochemical, state-of-the-art imaging, and recently developed gene editing methods to address the following questions: (1) What is the Ub- dependent mechanism that mediates crosstalk between mitochondrial fission and fusion machineries, and how does it maintain balance between these two processes? (2) What is the mechanism by which Rnf179 in concert with Mff and Drp1 controls mitochondrial homeostasis? (3) What is the mechanism by which Rnf179 in concert with Mff and Drp1 complex controls mitophagy?

概括线粒体位于与年龄有关的人类疾病的中心，例如帕金森氏症，阿尔茨海默氏症，亨廷顿疾病。随着当前人口老龄化的趋势，为这些疾病开发疗法是至关重要的需要。保护策略主要旨在中和有毒活性氧（ROS）部分成功。这是需要采用独立行动方式的新的治疗选择来改善当前疗法的成功率。拟议的工作直接应对这一需求，并将提供关键深入了解线粒体质量控制途径的调节和功能。真核细胞具有多层系统专用于线粒体质量控制。最初的防御由一个网络组成降低蛋白质的蛋白水解系统，这些蛋白质被损坏，折叠或倒置的蛋白质或控制活动压力响应性线粒体因素。最近的发现，包括PI实验室的工作，证明了通过依赖于UB的降解/控制的泛素（UB）/蛋白酶体系统（UPS）（UPS）相关蛋白对于线粒体质量控制至关重要。线粒体融合和裂变的调节（线粒体动力学）是另一种主要线粒体质量控制机制。最近我们提出了线粒体膜（OMM）蛋白的稳定性，包括线粒体融合因子MFN1和 MFN2由线粒体裂变蛋白DRP1和MFF控制。与此相一致，我们的新数据表明此过程需要先前未经跨度的UB依赖性信号传导函数。我们还发现，线粒体蛋白泛素化是局灶性的，并且在MFF - / - 加速了该过程。 DRP1 - / - 细胞。这些和其他初步结果表明，调节和协调线粒体质量控制途径，包括新的线粒体蛋白的参与线粒体动力学与线粒体之间的降解中间体和相互依赖。我们也是确定了与OMM相关的E3 UB连接酶RNF179，根据我们的初步发现控制包括MFN1和MFN2在内的OMM蛋白的稳定性，但也会影响线粒体特异性自噬（线粒体），这是一种关键的UPS依赖性线粒体质量控制途径。该提议的主要目标是测试以下假设：通过依赖于UB的信号传导，DRP1和MFF控制并协调不同的假设线粒体质量途径，包括线粒体裂变和融合速率以及线粒体的激活。我们还期望用线粒体蛋白泛素化定义RNF179的作用和机制线粒体功能的控制和线粒体质量控制途径的整合，包括parkin- 依赖和非依赖的线粒体。为了实现这些目标，我们结合了生化，最先进的成像，并且最近开发了基因编辑方法来解决以下问题：（1）UB-是什么介导线粒体裂变和融合机之间串扰的依赖机制，以及如何它在这两个过程之间保持平衡吗？（2）RNF179在与MFF和DRP1控制线粒体稳态的音乐会？（3）RNF179在与MFF和DRP1复合物控制Mitophagy的音乐会？