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?

概括线粒体是与年龄相关的人类疾病的核心，例如帕金森病、阿尔茨海默病、亨廷顿舞蹈病随着当前人口老龄化的趋势，开发这些疾病的治疗方法至关重要。保护策略主要旨在中和有毒的活性氧（ROS）。因此，需要具有独立作用模式的新治疗方案来改善。当前疗法的成功率。拟议的工作直接响应了这一需求，并将提供关键的信息。对真核细胞线粒体质量控制途径的调节和功能的深入了解。致力于线粒体质量控制的多层系统由一个网络组成。蛋白水解系统降解受损、错误折叠或错误定位的蛋白质，或控制最近的研究结果，包括 PI 实验室的工作，证明了应激反应线粒体因素。泛素 (Ub)/蛋白酶体系统 (UPS)，通过 OMM 的 Ub 依赖性降解/控制相关蛋白对于线粒体融合和裂变的质量控制至关重要。（线粒体动力学）是最近我们提出的另一个主要线粒体质量控制机制。线粒体外膜 (OMM) 蛋白的稳定性，包括线粒体融合因子 Mfn1 和我们的新数据表明，Mfn2 受线粒体裂变蛋白 Drp1 和 Mff 控制。这一过程需要以前未曾怀疑过的 Drp1 和 Mff 的 Ub 依赖性信号传导功能。我们还发现线粒体蛋白泛素化是焦点，并且该过程在 Mff-/- 和这些和其他初步结果表明，Drp1-/- 细胞的调节机制尚未得到重视。协调线粒体质量控制途径，包括新型线粒体蛋白的参与我们还研究了降解中间体以及线粒体动力学和线粒体自噬之间的相互依赖性。鉴定了 OMM 相关的 E3 Ub 连接酶 Rnf179，根据我们的初步发现，它不仅 OMM 蛋白（包括 Mfn1 和 Mfn2）的稳定性控制，但也会影响线粒体特异性自噬（线粒体自噬），一种关键的 UPS 依赖性线粒体质量控制途径该提案的主要目标。目的是检验以下假设：通过 Ub 依赖性信号传导，Drp1 和 Mff 控制和协调不同的线粒体质量途径，包括线粒体裂变和融合率以及线粒体自噬的激活。我们还期望通过线粒体蛋白泛素化来定义 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- 为了实现这些目标，我们结合了最先进的生化技术。成像，以及最近开发的基因编辑方法来解决以下问题：（1）什么是Ub- 介导线粒体裂变和融合机制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 的作用机制是什么？ (3) Rnf179 与 Mff 和 Drp1 共同控制线粒体稳态的机制是什么？ Mff 和 Drp1 复合物共同控制线粒体自噬？

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Outer mitochondrial membrane E3 Ub ligase MARCH5 controls mitochondrial steps in peroxisome biogenesis.

线粒体外膜 E3 Ub 连接酶 MARCH5 控制过氧化物酶体生物合成中的线粒体步骤。

DOI：
发表时间：
2023-09-01
期刊：
影响因子：
0
作者：
Verhoeven, Nicolas;Oshima, Yumiko;Cartier, Etienne;Neutzner, Albert;Boyman, Liron;Karbowski, Mariusz
通讯作者：
Karbowski, Mariusz

The OMM-severed and IMM-ubiquitinated mitochondria are intermediates of mitochondrial proteotoxicity-induced autophagy in PRKN/parkin-deficient cells.

OMM 切断和 IMM 泛素化线粒体是 PRKN/parkin 缺陷细胞中线粒体蛋白毒性诱导自噬的中间体。