In vivo Analysis of ATM-Regulated Pathways

ATM 调节通路的体内分析

• 批准号: 7915858
• 负责人: Randal Scot Tibbetts
• 金额: $ 10万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7915858
• 关键词: ATM gene; ATM wt Allele; Apoptosis; Ataxia Telangiectasia; Attenuated; Binding Proteins; Biochemical; CHEK2 gene; CREB-binding protein; CSNK1A1 gene; Cell Cycle; Cell Cycle Checkpoint; Cell Cycle Regulation; Cells; Circadian Rhythm Pathway; Circadian Rhythms; Cues; Cyclic AMP-Responsive DNA-Binding Protein; DNA Binding; DNA Damage; DNA Repair; Gene Expression; Gene Targeting; Genetic; Genotoxic Stress; Glean; Goals; In Vitro; Laboratories; Link; Malignant Neoplasms; Mammalian Cell; Mediating; Mediator of activation protein; Metabolism; Modeling; Modification; Molecular; Mutate; Mutation; Nature; Nerve Degeneration; Neurons; Oncogene Proteins; Outcome; Paint; Pathway interactions; Phenotype; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphorylation Site; Phosphotransferases; Physiological; Physiology; Play; Predisposition; Protein Binding; Protein Kinase; Proteins; Regulation; Regulator Genes; Role; Serum; Signal Transduction; Signal Transduction Pathway; Site; Stimulus; Stress; Syndrome; Tertiary Protein Structure; Testing; Transcription Coactivator; Tumor Suppressor Proteins; Work; ataxia telangiectasia mutated protein; base; casein kinase; casein kinase I; cell growth; chromatin immunoprecipitation; human CREBBP protein; in vivo; insight; interest; novel; programs; promoter; protein complex; protein function; relating to nervous system; research study; response; transcription factor

DESCRIPTION (provided by applicant): The major objectives of this application are to: (i) functionally characterize a signal transduction pathway linking ATM (ataxia-telangiectasia-mutated) to the cyclic AMP response element-binding protein (CREB) transcription factor; and (ii) identify and functionally characterize novel ATM substrates. ATM is a DNA damage-activated protein kinase that is mutated in the genetic instability and neurodegeneration syndrome, ataxia-telangiectasia, whereas CREB is a neuroprotective transcription factor that regulates cell growth, metabolism, and survival. We have defined a new mode of CREB regulation whereby ATM and casein kinases 1 and 2 (CK1/CK2) collaboratively phosphorylate CREB on five clustered sites termed the RAX domain (co-Regulated ATM and Casein Kinase Sites) in response to genotoxic stress. Phosphorylation of CREB by ATM and CK1/CK2 inhibits the interaction between CREB and its coactivator, CREB-binding protein (CBP) suggesting that the ATM pathway may repress CREB transcriptional functions in response to DNA damage. The linkage between ATM and CREB is intriguing given the neuroprotective functions of both factors. In this proposal we will test the hypothesis that ATM plays a dual role in CREB regulation through suppression and stimulation of RAX domain phosphorylation in unperturbed and DNA-damaged cells, respectively. An important goal of the work is to define the upstream signals controlling CREB RAX domain phosphorylation in the absence of DNA damage and to elucidate the biochemical outcomes of its modification in intact cells. In addition, we will use information gleaned from the CREB phosphorylation paradigm to discover and functionally characterize protein substrates that are coordinately regulated by ATM and CK1/CK2 in response to DNA damage. These studies should yield fundamental insights into the mechanisms of ATM function and CREB regulation, and may alter current views of ATM signaling in response to DNA damage. The goal of this project is to understand the molecular basis for the neurodegeneration/cancer susceptibility syndrome, ataxia-telangiectasia (A-T), which is caused by mutations in the ATM gene. ATM is a critical regulator of cellular responses to DNA damage and the work proposed in this application will characterize a particularly important downstream target of ATM, termed CREB (cyclic AMP response element-binding protein), which is an important regulator of gene expression. We are specifically interested in examining whether deregulation of CREB contributes to the manifestation of A-T-related phenotypes, including cancer, and neuron demise.

描述（由申请人提供）：本申请的主要目的是：（i）功能性表征连接ATM（共济失调-毛细血管扩张-突变）与环AMP反应元件结合蛋白（CREB）转录因子的信号转导途径;和（ii）鉴定和功能性表征新型ATM底物。ATM是一种DNA损伤激活的蛋白激酶，在遗传不稳定性和神经变性综合征，共济失调-毛细血管扩张症中突变，而CREB是一种调节细胞生长，代谢和存活的神经保护性转录因子。我们已经定义了一种新的CREB调节模式，即ATM和酪蛋白激酶1和2（CK 1/CK 2）协同磷酸化CREB的5个集群网站称为RAX域（共调节ATM和酪蛋白激酶网站），以响应遗传毒性应激。通过ATM和CK 1/CK 2磷酸化CREB抑制CREB与其辅激活因子CREB结合蛋白（CBP）之间的相互作用，表明ATM途径可能抑制CREB响应DNA损伤的转录功能。ATM和CREB之间的联系是有趣的，因为这两种因子都具有神经保护功能。在这个建议中，我们将测试的假设，ATM通过抑制和刺激RAX结构域的磷酸化在未受干扰和DNA损伤的细胞，分别在CREB调控中发挥双重作用。这项工作的一个重要目标是确定在没有DNA损伤的情况下控制CREB RAX结构域磷酸化的上游信号，并阐明其在完整细胞中修饰的生化结果。此外，我们将使用从CREB磷酸化范式收集的信息来发现和功能表征由ATM和CK 1/CK 2协调调节的蛋白质底物，以响应DNA损伤。这些研究将对ATM功能和CREB调控机制产生根本性的认识，并可能改变目前对ATM信号对DNA损伤的反应。该项目的目标是了解神经变性/癌症易感性综合征，共济失调-毛细血管扩张症（A-T）的分子基础，这是由ATM基因突变引起的。ATM是对DNA损伤的细胞反应的关键调节剂，并且在本申请中提出的工作将表征ATM的特别重要的下游靶标，称为CREB（环AMP反应元件结合蛋白），其是基因表达的重要调节剂。我们特别感兴趣的是研究CREB的失调是否有助于A-T相关表型的表现，包括癌症和神经元死亡。