Signaling mechanisms that detect stress and maintain homeostasis

检测压力和维持体内平衡的信号机制

基本信息

批准号：
10219290
负责人：
T Keith Blackwell
金额：
$ 49.62万
依托单位：
JOSLIN DIABETES CENTER
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-20 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10219290
关键词：
70-kDa Ribosomal Protein S6 Kinases Acute Affect Aging Area CRISPR/Cas technology Caenorhabditis elegans Cell physiology Data Disease Drug Metabolic Detoxication Enzyme Activation Event Growth Health Homeostasis Human Human Development Longevity Mass Spectrum Analysis Mediating Metabolic Mitochondria Modeling Modification Molecular Chaperones Nox enzyme Organism Orthologous Gene Oxidation-Reduction Phenotype Phosphotransferases Play Proteins Proto-Oncogene Proteins c-akt ROCK1 gene Reactive Oxygen Species Regulation Research Role Screening Result Signal Transduction Stress Work base biological adaptation to stress genome editing in vivo interest p38 Mitogen Activated Protein Kinase response sensor small molecule stress reactivity transcription factor

70-kDa Ribosomal Protein S6 Kinases Acute Affect Aging Area CRISPR/Cas technology Caenorhabditis elegans Cell physiology Data Disease Drug Metabolic Detoxication Enzyme Activation Event Growth Health Homeostasis Human Human Development Longevity Mass Spectrum Analysis Mediating Metabolic Mitochondria Modeling Modification Molecular Chaperones Nox enzyme Organism Orthologous Gene Oxidation-Reduction Phenotype Phosphotransferases Play Proteins Proto-Oncogene Proteins c-akt ROCK1 gene Reactive Oxygen Species Regulation Research Role Screening Result Signal Transduction Stress Work base biological adaptation to stress genome editing in vivo interest p38 Mitogen Activated Protein Kinase response sensor small molecule stress reactivity transcription factor

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项目摘要

Project Summary This MIRA proposal focuses on two overlapping areas: stress response regulation and the functions of redox-based signaling in vivo. It is a fundamentally important problem how organisms detect and respond to different forms of stress. Much has been learned in this area but we still have a very incomplete understanding of how some stresses are detected, including reactive small molecules such as ROS. For many years my group has studied stress responses and aging in C. elegans, focusing on the Nrf2 transcription factor ortholog SKN-1. Nrf2 mediates a conserved detoxification response to reactive small molecules but has many additional functions, and is of great importance in health and disease. Working in C. elegans we have defined a number of aspects of SKN-1/Nrf2 regulation and functions, including its major role in longevity assurance. We have recently uncovered an exciting mechanism of SKN-1/Nrf regulation that forms the basis for this new research direction. We find that SKN-1 and human Nrf2 are activated at the ER by a localized ROS signal that can derive from the ER, NOX enzyme activation induced by stress, or mitochondria. This signal induces sulfenylation of a single Cys within the kinase activation loop of the ER unfolded protein sensor IRE-1, resulting in acute inhibition of the IRE-1 unfolded protein response and activation of p38 signaling at IRE-1 through a second sulfenylation event. p38 signaling in turn activates SKN-1/Nrf2. Remarkably, other kinases of major interest (AKT, p70S6K, ROCK1) seem to be regulated through sulfenylation of the same Cys. The data reveal an unexpected IRE-1 function that is regulated by a redox switch, a major stress sensor for SKN- 1/Nrf2, and a possible rationale for how redox stress can affect so many cellular processes. They also suggest that the scope and functional versatility of Cys-based signaling are much wider than is generally appreciated. In our proposed research we will continue to identify mechanisms of SKN-1/Nrf2 regulation and their functions in vivo, but will also cast our net wider in utilizing the advantages of C. elegans to explore mechanisms and functions of Cys redox signaling in the context of stress, growth, and other conditions. We will refine models for IRE-1 regulation of SKN-1/Nrf2 and its functions in vivo. We will also similarly study SKN- 1/Nrf2 regulation by the chaperone TRIC, another mechanism we have identified that may involve redox signaling, and build upon screening results to develop new models for SKN-1/Nrf2 regulation. Using mass spectrometry (MS), we will collaboratively identify C. elegans proteins that are Cys-sulfenylated under stress and growth conditions. We will investigate regulatory and in vivo implications of this modification for the kinases indicated above, and candidates chosen from our MS data. C. elegans will be ideal for this work because of the relative rapidity of Cas9/CRISPR genome editing, and phenotypic analyses. Our research will reveal stress-responsive regulatory mechanisms of fundamental interest, and take major steps towards identifying new mechanistic targets and functional implications of redox signaling in vivo.

项目摘要 MIRA提案重点介绍两个重叠领域：压力反应调节和功能基于氧化还原的信号在体内。这是一个根本重要的问题，生物如何检测和回应不同形式的压力。在这一领域已经学到了很多东西，但我们仍然有一个不完整的理解如何检测到一些应力，包括反应性的小分子，例如ROS。多年来我小组研究了秀丽隐杆线虫的压力反应和衰老，重点是NRF2转录因子直系同源 SKN-1。 NRF2介导了对反应性小分子的保守排毒响应，但具有许多其他功能，在健康和疾病中非常重要。在秀丽隐杆线虫中工作，我们定义了 SKN-1/NRF2调节和功能的许多方面，包括其在寿命保证中的主要作用。我们最近发现了SKN-1/NRF调节的令人兴奋的机制，这是构成的基础这个新的研究方向。我们发现SKN-1和人NRF2通过局部ROS在ER处激活可以源自ER的信号，由应力或线粒体引起的NOX酶激活。这个信号在ER展开的蛋白传感器IRE-1，导致急性抑制IRE-1展开的蛋白质反应和IRE-1信号的激活通过第二次磺酰化事件。 p38信号依次激活SKN-1/NRF2。值得注意的是，其他激酶主要感兴趣（AKT，P70S6K，ROCK1）似乎通过同一CYS的磺酰化来调节。这数据揭示了由氧化还原开关调节的意外IRE-1函数，氧化还原开关是SKN-的主要应力传感器 1/NRF2，以及氧化还原应力如何影响许多细胞过程的可能理由。他们也建议基于CYS的信号传导的范围和功能多功能性远远超过通常所欣赏的范围。在我们提出的研究中，我们将继续确定SKN-1/NRF2调节的机制及其在体内功能，但也会在利用秀丽隐杆线虫的优势探索方面更广泛在压力，生长和其他条件下，CYS氧化还原信号的机制和功能。我们将完善用于SKN-1/NRF2的IRE-1调节模型及其在体内的功能。我们还将同样研究skn- 1/NRF2由伴侣Tric调节，我们已经确定的另一种可能涉及氧化还原的机制信号传导，并建立在筛选结果的基础上，以开发SKN-1/NRF2调节的新模型。使用质量光谱法（MS），我们将协作识别秀丽隐杆线虫蛋白，这些秀丽隐杆菌在应激下是硫二烯基的和生长条件。我们将调查这种修饰的调节和体内含义上面指示的激酶以及从我们的MS数据中选择的候选者。秀丽隐杆线虫将是这项工作的理想选择由于CAS9/CRISPR基因组编辑和表型分析的相对速度。我们的研究会揭示基本兴趣的压力响应性调节机制，并采取重大步骤识别体内氧化还原信号传导的新机械目标和功能含义。