In vivo Analysis of ATM-Regulated Pathways

ATM 调节通路的体内分析

基本信息

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
项目类别：
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

项目摘要

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）：本申请的主要目标是：（i）功能性地表征连接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； (ii) 识别新型 ATM 底物并对其进行功能表征。 ATM 是一种 DNA 损伤激活蛋白激酶，在遗传不稳定和神经变性综合征、共济失调毛细血管扩张症中发生突变，而 CREB 是一种神经保护性转录因子，可调节细胞生长、代谢和存活。我们定义了一种新的 CREB 调节模式，其中 ATM 和酪蛋白激酶 1 和 2 (CK1/CK2) 在称为 RAX 结构域（共同调节的 ATM 和酪蛋白激酶位点）的五个聚集位点上协同磷酸化 CREB，以响应基因毒性应激。 ATM 和 CK1/CK2 对 CREB 的磷酸化会抑制 CREB 与其共激活剂 CREB 结合蛋白 (CBP) 之间的相互作用，表明 ATM 途径可能会抑制 CREB 转录功能以应对 DNA 损伤。鉴于 ATM 和 CREB 都具有神经保护功能，两者之间的联系很有趣。在本提案中，我们将测试以下假设：ATM 通过分别抑制和刺激未受干扰的细胞和 DNA 受损细胞中的 RAX 结构域磷酸化，在 CREB 调节中发挥双重作用。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，并阐明其修饰在完整细胞中的生化结果。此外，我们将利用从 CREB 磷酸化范例中收集的信息来发现和功能表征由 ATM 和 CK1/CK2 协调调节以响应 DNA 损伤的蛋白质底物。这些研究应该对 ATM 功能和 CREB 调节机制产生基本的见解，并可能改变目前对 ATM 信号响应 DNA 损伤的看法。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，该综合征是由 ATM 基因突变引起的。 ATM 是细胞对 DNA 损伤反应的关键调节因子，本申请中提出的工作将表征 ATM 的一个特别重要的下游靶标，称为 CREB（环 AMP 反应元件结合蛋白），它是基因表达的重要调节因子。我们特别感兴趣的是检查 CREB 的失调是否会导致 A-T 相关表型的表现，包括癌症和神经元死亡。