Roles and Regulation of the bZip Transcription Factor CEBP-1 in Stress Response Pathways

bZip 转录因子 CEBP-1 在应激反应途径中的作用和调节

• 批准号: 9897413
• 负责人: Rose Malinow
• 金额: $ 3.84万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9897413
• 关键词: Affect; Animal Model; Animals; Binding; Binding Proteins; Biological Assay; Biological Process; CCAAT-Enhancer-Binding Protein-alpha; CCAAT-Enhancer-Binding Proteins; Caenorhabditis elegans; Cardiovascular Diseases; Cell Cycle Regulation; Cell physiology; Cells; Cellular Stress Response; Cytoskeleton; DNA Binding; DNA Binding Domain; DNA-Binding Proteins; Defect; Development; Disease; Dissection; Gene Expression; Genes; Genetic; Genetic Screening; Genetic Transcription; Goals; Hematopoiesis; Heterodimerization; Homologous Protein; Immune response; Immunoprecipitation; Lead; Leucine Zippers; Link; MAP Kinase Gene; MAP Kinase Kinase Kinase; MAP Kinase Modules; MAPK1 gene; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Microtubules; Molecular; Mutation; N-terminal; Neurodegenerative Disorders; Neurons; Output; Pathway interactions; Phenotype; Phosphotransferases; Play; Post-Translational Protein Processing; Process; Promoter Regions; Protein Family; Proteins; Regulation; Reporter; Role; Signal Pathway; Signal Transduction; Skeletal Development; Specificity; Stimulus; Stress; Synapses; System; Testing; Transactivation; Transcriptional Regulation; Transgenic Organisms; axon injury; axon regeneration; bZIP Domain; base; biological adaptation to stress; causal variant; cell type; genome editing; human disease; in vivo; insight; interest; loss of function; new therapeutic target; novel; null mutation; p38 Mitogen Activated Protein Kinase; promoter; protein function; protein protein interaction; repaired; response; spatiotemporal; transcription factor

Project Summary Proper cellular response to stress is vital for organismal survival. Most cellular stress responses involve the action of transcription factors that regulate expression of stress response genes. Determining how transcription factors can initiate differential responses to a variety of stressful inputs is critical for understanding multiple human diseases. The overall goal of this project is to uncover the mechanisms by which a single transcription factor can integrate multiple inputs and direct proper transcriptional responses. CCAAT enhancer binding proteins (C/EBP) family proteins are involved in many forms of stress responses. C/EBPs contain a basic-leucine-zipper DNA binding domain, and transactivating and regulatory domains, which modulate transcriptional activity via specific protein-protein interactions or post-translational modifications. Although C/EBP proteins have been extensively studied, the mechanism of their action under external stimuli is still poorly understood. Studies using Caenorhabditis elegans offer great advantages to investigate multi-level cellular and organismal responses to stress signals. This proposal focuses on examining the molecular function of the conserved C/EBP homologue, CEBP-1, in mediating both development and neuronal stress responses. In neurons, CEBP-1 plays critical roles in response to axon injury and cytoskeleton disruption, acting downstream of the DLK-1 MAP kinase cascade. In animal development CEBP-1 is negatively controlled by the Tribbles family protein, NIPI-3, and in turn activates a different MAP kinase cascade. Using powerful forward genetic screening, I have identified a distinct functional domain in the N-terminus of CEBP-1 that is crucial for its activity. Here, I propose to test the hypothesis that this domain, in conjunction with differential protein-protein interaction, endows transactivating regulation to initiate transcriptional responses to different stressful inputs. The goals of this study will be accomplished through the following specific aims: Aim 1: To identify CEBP-1 binding proteins using IP- mass spectrometry and to demonstrate that the N-terminal region contains transactivating activity. Aim 2: To determine the mechanism of transcriptional regulation of CEBP-1 by NIPI-3; and Aim 3: to Identify new genetic players in NIPi-3/Tribbles and CEBP-1 pathway. The completion of this proposal will provide a deeper understanding of the general process of stress response to various external stimuli. Understanding the mechanism of this control could allow for development of targeted treatments for diseases affecting cellular stress response and repair.

项目摘要 细胞对压力的适当反应对生物体的生存至关重要。大多数细胞应激反应涉及 调节应激反应基因表达的转录因子的作用。确定转录如何 因素可以引发对各种压力输入的不同反应，这对于理解多重压力输入至关重要。 人类疾病。这个项目的总体目标是揭示一个单一的 转录因子可以整合多个输入并指导适当的转录反应。CCAAT 增强子结合蛋白（C/EBP）家族蛋白参与许多形式的应激反应。C/EBPs 含有碱性亮氨酸拉链DNA结合结构域，以及反式激活和调节结构域，其调节 通过特异性蛋白质-蛋白质相互作用或翻译后修饰来调节转录活性。虽然 C/EBP蛋白已被广泛研究，但其在外界刺激下的作用机制尚不清楚 明白使用秀丽隐杆线虫的研究为研究多水平细胞和 对压力信号的反应。这项建议的重点是检查的分子功能， 保守的C/EBP同源物，CEBP-1，在介导发育和神经元应激反应。在 神经元，CEBP-1在响应轴突损伤和细胞骨架破坏中起关键作用，作用于下游 DLK-1 MAP激酶级联反应。在动物发育中，CEBP-1由Tribbles家族负控制 蛋白质NIPI-3，并反过来激活不同的MAP激酶级联。利用强大的前向遗传筛选， 我已经确定了一个独特的功能域的N-末端的CEBP-1是至关重要的，其活动。这里我 我建议测试这一假设，该域，结合差异蛋白质-蛋白质相互作用，赋予 反式激活调节以启动对不同应激输入的转录应答。本研究的目标 将通过以下具体目标来实现：目标1：使用IP-1鉴定CEBP-1结合蛋白， 质谱分析并证明N-末端区域含有反式激活活性。目标2： 目的3：通过NIPI-3对CEBP-1基因的转录调控机制的研究， NIPi-3/Tribbles和CEBP-1途径的参与者。这项提案的完成将提供一个更深入的 了解对各种外部刺激的应激反应的一般过程。了解 这种控制机制可以允许开发针对影响细胞的疾病的靶向治疗， 应激反应和修复。