Physiological role of the TNKS-PI31 dependent proteasome regulation

TNKS-PI31 依赖性蛋白酶体调节的生理作用

• 批准号: 9406701
• 负责人: Pak Cho
• 金额: $ 20.8万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9406701
• 关键词: 26S proteasome; ADP Ribose Transferases; ADP ribosylation; Affect; Affinity; Alzheimer' s Disease; Animal Experiments; Antibodies; Attenuated; Award; Binding; Biochemistry; Biological; Biological Process; Biology; Biophysics; Bortezomib; Cell Cycle Regulation; Cell Line; Cells; Cellular biology; Centromere; Clinical; Colon Adenocarcinoma; Colorectal Adenocarcinoma; Complex; Computer Simulation; Data; Defect; Disease; Disease model; Dissection; Drosophila genus; Drug Targeting; Future; Genetic; Genetic study; Goals; Growth; Homeostasis; Human; Huntington Disease; Knockout Mice; Link; Malignant Neoplasms; Mediating; Methods; Modeling; Molecular Chaperones; Multiple Myeloma; Mus; Neurodegenerative Disorders; Orthologous Gene; Parkinson Disease; Pathway interactions; Peptide Hydrolases; Pharmacology; Phase; Physiological; Play; Process; Proteasome Binding; Proteasome Inhibitor; Proteins; Proteomics; Regulation; Regulatory Pathway; Resolution; Role; Secondary to; Stress; System; Tankyrase; Telomere Maintenance; Testing; Therapeutic Uses; Time; Translational Research; Ubiquitin; age related; cancer therapy; experimental study; extracellular; fly; human disease; in vivo; inhibitor/antagonist; insight; interdisciplinary approach; multicatalytic endopeptidase complex; mutant; novel; protein aggregate; protein degradation; proteostasis; proteotoxicity; public health relevance; screening; small molecule; small molecule inhibitor; structural biology; tool

 DESCRIPTION (provided by applicant): Physiological role of the TNKS-PI31 dependent proteasome regulation Protein degradation by the ubiquitin-proteasome system is central to cell homeostasis and survival. Defects in this process are associated with many human diseases, including cancer and neurodegenerative disorders. The 26S proteasome is a large protease complex that degrades ubiquitinated proteins. We recently showed that ADP-ribosylation promotes 26S proteasome activity in both Drosophila and human cells. We identified the ADP-ribosyltransferase Tankyrase (TNKS) and the 19S assembly chaperones as direct binding partners of the proteasome regulator PI31 (Proteasome Inhibitor of 31kDA). TNKS-mediated ADP-ribosylation of PI31 promotes 26S assembly by modifying its affinity for proteasome proteins. Inhibition of TNKS by a small-molecule inhibitor, XAV939, blocks this process - revealing a novel mechanism of proteasome regulation that can be targeted with existing small-molecule inhibitors - as well as attenuating the growth of multiple myeloma cells that are particularly sensitive to fluctuations in proteasome activity. These results suggest a potential new link between NAD+, formation of toxic protein aggregates, cancer growth and proteasome regulation that likely play a central role in protein homeostasis. Here, we propose aims to further dissect this regulatory mechanism. Specifically, during the K99/R00 phase of the award, we intend to characterize the newly identified Archipelago (Ago; Drosophila ortholog of Fbw7)-PI31 interaction and develop tools to understand the physiological role of the TNKS- dependent proteasome activation. Furthermore, by exploiting the TNKS-PI31 pathway, we will be able to rigorously explore the translational research potential of proteasome in various human disorders that arise secondary to dysfunctional protein homeostasis. For these purposes, we will use a multi-disciplinary approach that integrates genetics, cell biology, biochemistry and structural biology in the context of human disease models in Drosophila and mice, as well as in human multiple myeloma and colon adenocarcinoma cell lines. We expect that this project will fundamentally advance our understanding of how protein degradation is regulated and provide new insights into how we can manipulate this process to treat human diseases.

 描述（由申请人提供）：TNKS-PI31 依赖性蛋白酶体调节的生理作用泛素蛋白酶体系统的蛋白质降解对于细胞稳态和存活至关重要，该过程中的缺陷与许多人类疾病有关，包括癌症和神经退行性疾病。 26S 蛋白酶体是一种大型蛋白酶复合物，可降解泛素化蛋白质，我们最近表明 ADP-核糖基化可促进。我们确定了果蝇和人类细胞中的 26S 蛋白酶体活性，ADP-核糖基转移酶端聚合酶 (TNKS) 和 19S 组装伴侣是蛋白酶体调节剂 PI31（TNKS 介导的 PI31 的 ADP 核糖基化的蛋白酶体抑制剂）的直接结合伴侣。通过修改其对蛋白酶体蛋白的亲和力进行 26S 组装。小分子抑制剂 XAV939 对 TNKS 的抑制可阻断这一过程，揭示了一种新的蛋白酶体调节机制，可以用现有的小分子抑制剂来靶向，并减弱对波动特别敏感的多发性骨髓瘤细胞的生长这些结果表明 NAD+、有毒蛋白质聚集体的形成、癌症生长和蛋白酶体调节之间存在潜在的新联系，这可能在蛋白质稳态中发挥核心作用。旨在进一步提出 具体来说，在该奖项的 K99/R00 阶段，我们打算表征新发现的群岛（Ago；Fbw7 的果蝇直系同源物）-PI31 相互作用，并开发工具来了解 TNKS 依赖性蛋白酶体的生理作用。此外，通过利用 TNKS-PI31 通路，我们将能够严格探索蛋白酶体在各种继发性人类疾病中的转化研究潜力。为了这些目的，我们将在果蝇和小鼠以及人类多发性骨髓瘤和结肠腺癌的人类疾病模型的背景下，使用一种多学科方法，将遗传学、细胞生物学、生物化学和结构生物学结合起来。我们期望这个项目将从根本上增进我们对蛋白质降解如何调节的理解，并为我们如何操纵这一过程来治疗人类疾病提供新的见解。