Molecular Chaperones and Protein Degradation

分子伴侣和蛋白质降解

基本信息

批准号：
9912181
负责人：
ALFRED L GOLDBERG
金额：
$ 54.58万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
1995
资助国家：
美国
起止时间：
1995-08-01 至 2021-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9912181
关键词：
26S proteasome Affect Alzheimer&apos s Disease Attention Binding Binding Proteins Biochemical Biological Brain Cell Cycle Progression Cells Clustered Regularly Interspaced Short Palindromic Repeats Complex Cryoelectron Microscopy Cyclic AMP Cyclic AMP-Dependent Protein Kinases Cyclic GMP-Dependent Protein Kinases Enzymes Epinephrine Exercise Fasting Genes Glucagon Goals Heart Heat-Shock Response Hepatocyte Hormones Human Learning Liver Mammalian Cell Mediating Modification Molecular Chaperones Mus Muscle Muscular Atrophy Mutation Neurodegenerative Disorders Neurons Neurotransmitters Normal Cell Pathway interactions Phosphorylation Phosphotransferases Physiological Production Proteasome Binding Protein Kinase Proteins Proteolysis Proteomics Quality Control RAD23B gene Regulation Reporting Strenuous Exercise Stress Structure Testing Therapeutic Tissues Ubiquitin Ubiquitination calmodulin-dependent protein kinase II in vivo interest misfolded protein multicatalytic endopeptidase complex mutant novel parkin gene/protein particle protein activation protein degradation proteostasis proteotoxicity tool ubiquilin

项目摘要

Project Summary / Abstract The great majority of proteins in mammalian cells are degraded by the ubiquitin (Ub) proteasome pathway (UPS). Although it is generally assumed that rates of protein breakdown by the UPS are determined solely through control of ubiquitination, recent studies have shown that the proteolytic capacity of 26S proteasomes is also tightly regulated and influences rates of proteolysis in cells. We recently made the exciting finding that protein kinase A (PKA) by phosphorylating subunit RPN6 enhances the proteasome’s multiple activities and thereby increases the cell’s capacity to degrade misfolded, aggregation-prone proteins, including mutant proteins that cause Alzheimer’s Disease and ALS. Because of its importance in cell regulation and therapeutic potential, we are pursuing in depth studies of the biochemical mechanisms and physiological importance of 26S phosphorylation by PKA and other protein kinases, as well as studies of the newly discovered ability of several proteasome-binding proteins to stimulate its activities. We hope to understand more fully how PKA enhances proteasome function and influences the structure of its 19S regulatory particle. Proteomic studies are planned to identify the short-lived cell proteins degraded faster upon Rpn6 phosphorylation. A valuable tool in these biochemical and structural studies will be construction by CRISPR gene editing of mutant lines carrying phosphomimetic and phosphodead Rpn6 mutations. Selected studies will test if three other kinases reported to phosphorylate proteasome subunits (e.g. protein Kinase G, CamKinase II, and DYRK2), or others alter proteasome function and protein turnover in similar ways as PKA. cAMP/PKA mediate the actions of many hormones and neurotransmitters, and we recently showed that epinephrine and glucagon trigger proteasome activation in hepatocytes by this mechanism. Upon fasting of mice, in muscle and liver Rpn6 becomes phosphorylated and proteasomes activated, as we also found in human muscles after intense exercise. We plan to explore further these actions, which are of clear physiological interest and demonstrate that surprisingly many major hormones can rapidly enhance protein homeostasis by altering proteasome function. Related studies will probe the mechanisms of proteasome activation by certain 26S-binding proteins. 1) The ZFAND protein, ZNF216, which is induced and essential for muscle atrophy, stimulates the proteasome’s degradative activity. 2) The related ZFAND protein, AIRAP, which is induced in heat shock, may cause the marked activation of 26S proteasomes that we recently discovered occurs rapidly on heat shock. 3) We also recently found that the UBL domain, through which many proteins bind to the 26S, by itself can stimulate proteasome activity. We believe this activation is an important new aspect of the functioning of the DUB Usp14 and of the UBL-UBA shuttling factors (e.g. Rad23) that deliver Ub conjugates to the proteasome.

项目摘要 /摘要哺乳动物细胞中绝大多数蛋白质被泛素（UB）蛋白酶体降解途径（UPS）。尽管通常假定确定UPS的蛋白质分解速率仅通过控制泛素化，最近的研究表明26S的蛋白水解能力蛋白酶体也受到严格调节，并影响细胞中蛋白水解的速率。我们最近使令人兴奋的通过磷酸化亚基RPN6发现该蛋白激酶A（PKA）增强了蛋白酶体的多重活动，从而增加了细胞降解错误折叠的，可聚集的蛋白质的能力，包括引起阿尔茨海默氏病和ALS的突变蛋白。因为它在细胞调节中的重要性以及治疗潜力，我们正在深入研究生化机制和生理学 PKA和其他蛋白激酶的26S磷酸化的重要性，以及新的研究发现了几种蛋白酶体结合蛋白刺激其活性的能力。我们希望理解 PKA如何增强蛋白酶体功能并影响其19S调节粒子的结构。计划蛋白质组学研究以鉴定RPN6时更快降解的短寿命细胞蛋白磷酸化。这些生化和结构研究中的有价值的工具将是CRISPR的建设携带磷酸化和磷酸化RPN6突变的突变系的基因编辑。选定的研究将测试其他三个激酶是否报告给磷酸化蛋白酶体亚基（例如蛋白激酶G，凸轮激酶II，和dyrk2）或其他人以与PKA相似的方式改变了蛋白质更新。营地/PKA调解许多骑马和神经递质的动作，我们最近表明肾上腺素和胰高血糖素通过该机制触发肝细胞中的蛋白酶体激活。禁食在肌肉和肝脏中的小鼠rpn6被磷酸化并激活蛋白酶体，正如我们在强烈运动后的人类肌肉。我们计划进一步探索这些行动，这些行动很清楚生理兴趣，并证明令人惊讶的是许多主要激素会迅速增强蛋白质通过改变蛋白酶体功能来稳态。相关研究将探测蛋白酶体的机制某些26S结合蛋白的激活。 1）ZFAND蛋白ZnF216，该蛋白是诱导的，对于肌肉萎缩，刺激蛋白酶体的降解活性。 2）相关的ZFAND蛋白Airap，该蛋白是在热休克中诱导的，可能导致我们最近发现的26S蛋白酶体的明显激活迅速发生在热冲击上。 3）最近我们还发现，UBL域，许多蛋白质通过与26S结合，本身可以刺激蛋白酶体活性。我们认为这种激活是一个重要的新事物 DUB USP14和UBL-UBA穿梭因子（例如RAD23）功能的方面结合蛋白酶体。

项目成果

期刊论文数量（79）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Control of proteasomal proteolysis by mTOR.

DOI：
10.1038/nature16472
发表时间：
2016-01-21
期刊：
Nature
影响因子：
64.8
作者：
Zhao J;Garcia GA;Goldberg AL
通讯作者：
Goldberg AL

Raising cGMP restores proteasome function and myelination in mice with a proteotoxic neuropathy.

提高 cGMP 可恢复患有蛋白毒性神经病的小鼠的蛋白酶体功能和髓鞘形成。

DOI：
10.1093/brain/awab249
发表时间：
2022
期刊：
Brain : a journal of neurology
影响因子：
0
作者：
VerPlank,JordanJS;Gawron,Joseph;Silvestri,NicholasJ;Feltri,MLaura;Wrabetz,Lawrence;Goldberg,AlfredL
通讯作者：
Goldberg,AlfredL

Isolation of mammalian 26S proteasomes and p97/VCP complexes using the ubiquitin-like domain from HHR23B reveals novel proteasome-associated proteins.