Defining the roles of ubiquitination during the environmental stress response

定义泛素化在环境应激反应中的作用

• 批准号: 8951314
• 负责人: Gustavo M Silva
• 金额: $ 10.27万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8951314
• 关键词: Affect; Alzheimer' s Disease; Antioxidants; Award; Binding; Biochemical; Biological Assay; Cell Death; Cell Survival; Cell physiology; Cells; Co-Immunoprecipitations; Complex; Computational Biology; Computational Technique; Coupled; DNA; Data; Deubiquitinating Enzyme; Development; Dimensions; Disease; Disease Progression; Dissociation; Drug Targeting; Early Diagnosis; Environment; Environmental Risk Factor; Enzymes; Exposure to; Failure; Future; Genes; Genomics; Goals; Heating; Homeostasis; Investigation; Ionizing radiation; Lipids; Lysine; Mammalian Cell; Mammals; Maps; Mass Spectrum Analysis; Mediating; Mentors; Messenger RNA; Methodology; Modeling; Molecular; Molecular Biology; Molecular Genetics; Mus; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Nucleotides; Outcome; Oxidants; Oxidation-Reduction; Oxidative Stress; Parkinson Disease; Pathway interactions; Phase; Polyribosomes; Polyubiquitin; Polyubiquitination; Positioning Attribute; Post-Translational Protein Processing; Protein Biosynthesis; Proteins; Proteomics; Regulatory Pathway; Research; Resistance; Resolution; Ribosomal Proteins; Ribosomes; Role; Saccharomyces cerevisiae; Signal Pathway; Signal Transduction; Site; Specificity; Staging; Stress; System; Technology; Testing; Training; Translations; Ubiquitin; Ubiquitination; Yeasts; base; biological adaptation to stress; career; cofactor; design; genetic approach; genetic variant; genome-wide; innovation; insight; mRNA Expression; multicatalytic endopeptidase complex; multidisciplinary; mutant; novel; novel therapeutics; overexpression; pollutant; prevent; protein aggregate; protein expression; protein protein interaction; public health relevance; response; targeted treatment; therapy development; tool; treatment strategy; tumor; tumor progression

 DESCRIPTION (provided by applicant): The goal of this proposal is to investigate the mechanisms by which protein ubiquitination regulates cellular resistance to oxidative stress. Increased exposure to oxidants combined to the accumulation of toxic protein aggregates is the cause of several neurodegenerative diseases such as Parkinson's and Alzheimer's. Oxidative stress is a very prominent type of stress induced by exposure to diverse environmental factors such as ionizing radiation, heat, and pollutants. Oxidative stress damages several biomolecules including lipids, DNA and proteins, potentially resulting in cell death. To prevent cell death and disease progression, cells use ubiquitination as the signal for removing damaged proteins through the proteasome and thus preventing aggregation. However, many non-degradative roles for ubiquitination have been described which are signaled by different lysine (K) ubiquitin chains when conjugated to target proteins. Using yeast S. cerevisiae as a model, we found that the unconventional K63 polyubiquitin chain accumulates strongly and dynamically, and impacts cellular viability in response to oxidative stress. Using innovative proteomics approaches, we identified many ribosome proteins as targets of K63 ubiquitination. However, the mechanisms by which K63 ubiquitination regulates cellular viability are still unknown. Therefore, in Aim1 system-wide approaches will be employed to investigate how K63 ubiquitination affects protein expression (K99). In Aim 2, interaction analysis will be performed to define the mechanisms that regulate the specificity of the K63 ubiquitin pathway in response to oxidative stress (K99/R00). Further, it has been demonstrated that mammalian cells accumulate K63 ubiquitin in response to oxidative stress, but the evolutionary conservation of this redox pathway is completely unknown. In Aim 3, the redox K63 ubiquitin system will be taken to mammalian cells to investigate overall conservation by mapping this pathway to the stress response in neuronal cells (R00). The outcome of this project will unravel novel mechanisms to increase cell tolerance to stress depending on selective ubiquitination. My entire career focused on studying different regulatory aspects of the ubiquitin proteasome system in response to stress. During the K99 mentored phase of this award, I will obtain training in genomics and computational techniques that will enable the investigation of one more layer of cellular signaling mediated by ubiquitin. NYU offers an ideal combination of intellectual environment and multidisciplinary facilities for th development of the K99 phase of this project. My long-term career goal is to establish my independent research group that will combine systems-wide methodologies with molecular biology and genetic approaches to decode the ubiquitination signals in response to stress. This award offers a unique opportunity to achieve this goal by enabling precise training and career guidance from my mentors and committee for my transition to independence. Understanding the multiple ubiquitin roles in cellular response to stress and the specificity of each distinct pathwa will provide groundbreaking insights into innovative strategies for treatment and early diagnosis of many damage-related disorders.

 描述（由申请人提供）：本提案的目的是研究蛋白质泛素化调节细胞抗氧化应激的机制。暴露于氧化剂的增加结合有毒蛋白质聚集体的积累是几种神经退行性疾病如帕金森氏症和阿尔茨海默氏症的原因。氧化应激是由暴露于各种环境因素（如电离辐射、热和污染物）引起的非常突出的应激类型。氧化应激损伤几种生物分子，包括脂质、DNA和蛋白质，可能导致细胞死亡。为了防止细胞死亡和疾病进展，细胞使用泛素化作为通过蛋白酶体去除受损蛋白质的信号，从而防止聚集。然而，已经描述了泛素化的许多非降解作用，当与靶蛋白缀合时，其通过不同的赖氨酸（K）泛素链发出信号。利用酵母S.作为模型，我们发现非常规的K63多聚泛素链强烈且动态地积累，并影响细胞对氧化应激的反应。利用创新的蛋白质组学方法，我们确定了许多核糖体蛋白作为K63泛素化的靶点。然而，K63泛素化调节细胞活力的机制仍然未知。因此，在Aim 1中，将采用全系统方法来研究K63泛素化如何影响蛋白质表达（K99）。在目标2中，将进行相互作用分析，以确定调节K63泛素途径响应氧化应激（K99/R 00）的特异性的机制。此外，它已被证明，哺乳动物细胞积累K63泛素在氧化应激反应，但这种氧化还原途径的进化保守是完全未知的。在目标3中，氧化还原K63泛素系统将被带到哺乳动物细胞中，通过将该途径映射到神经元细胞中的应激反应来研究整体保守性（R 00）。该项目的结果将揭示新的机制，以增加细胞耐受压力依赖于选择性泛素化。我的整个职业生涯都集中在研究泛素蛋白酶体系统对压力的不同调节方面。在该奖项的K99指导阶段，我将获得基因组学和计算技术方面的培训，这将使泛素介导的细胞信号传导的另一层研究成为可能。纽约大学为该项目K99阶段的开发提供了智力环境和多学科设施的理想组合。我的长期职业目标是建立一个独立的研究小组，将联合收割机的全系统方法与分子生物学和遗传学方法相结合，以解码应激反应中的泛素化信号。这个奖项提供了一个独特的机会来实现这一目标，使我的导师和委员会为我过渡到独立提供精确的培训和职业指导。了解多种泛素在细胞应激反应中的作用以及每种不同途径的特异性，将为许多损伤相关疾病的治疗和早期诊断的创新策略提供突破性的见解。