Temporal coordination of stimulus-induced gene expression by RNA-binding proteins

RNA结合蛋白刺激诱导的基因表达的时间协调

• 批准号: 10671719
• 负责人: Neelanjan Mukherjee
• 金额: $ 38.88万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10671719
• 关键词: Adrenal Glands; Aldosterone; Angiotensin II; Attenuated; Binding; Binding Proteins; Biological Models; Blood Pressure; Cells; Cholesterol; Ensure; Gene Expression; Gene Expression Regulation; Genetic Transcription; Goals; Human; Kinetics; Ligands; Messenger RNA; Methods; Pattern; Peptides; Phenotype; Physiologic pulse; Play; Post-Transcriptional Regulation; Prevention; Production; RNA; RNA Decay; RNA Processing; RNA-Binding Proteins; Research; Resolution; Role; Steroid biosynthesis; Stimulus; System; Testing; Textbooks; Translations; ZFP36L2 gene; Zona Glomerulosa; cytokine; immune activation; innovation; insight; mRNA Decay; mRNA Expression; mRNA Translation; posttranscriptional; programs; receptor; response

SUMMARY Cellular responses to external stimuli require dynamic gene regulation to facilitate rapid activation and effective resolution. Although transcriptional responses are a textbook mechanism for stimulus-induced responses, post- transcriptional regulation of mRNA translation and decay are the other side of the coin and are essential for rapid and adaptable stimulus-induced responses. For example, immune activation generates a prototypical temporal expression pattern for cytokine mRNAs known as an impulse response, which is a rapid pulse-like increase and decrease in expression. The destabilization of cytokine mRNAs is required for proper resolution of the impulse response and prevention of excess cytokine production. It is increasingly recognized that RNA-binding proteins (RBPs) play critical roles in achieving an appropriate stimulus-induced gene expression response by regulating RNA processing, decay, and translation of target RNAs. The over-arching goal of our research program is to understand how RBPs temporally coordinate stimulus-induced gene expression and cellular phenotypes. The model system we will use to study how dynamic RBP-RNA regulatory interactions temporally coordinate stimulus-induced gene expression is human adrenal steroidogenesis. This is a classic ligand-induced system in which the small peptide Angiotensin II (AngII) binds to its receptor in adrenal zona glomerulosa cells to stimulate the production of aldosterone, the master regulator of blood pressure. Thus, tight control of regulatory timing is required since it must be rapidly produced de novo from cholesterol in response to AngII. We have recently demonstrated that AngII treatment of immortalized and primary adrenocortical cells results in an impulse response. Furthermore, we found specific RBPs and regulated RNA decay facilitates the rapid implementation and resolution of the AngII impulse response. In this proposal, we will test the hypothesis that MSI2 and ZFP36L2 are counteracting forces that, respectively, potentiate and attenuate the kinetics of stimulus-induced mRNA levels by modulating target mRNA decay and translation, to ensure the proper timing and amplitude of the impulse response. We will utilize cutting edge methods to gain temporal and quantitative insights into how stimulus-induced changes in RBP binding determine changes in target mRNA expression responses and ultimately cellular phenotype. The use of innovative approaches we use will help shift the field from a static to a dynamic view of RBP-mRNA interactions and achieve mechanistic insight on how regulatory RBP-mRNA interactions control mRNA fate decisions to govern cellular responses.

概括 细胞对外界刺激的反应需要动态的基因调控，以促进快速激活和有效 解决。尽管转录反应是刺激诱导反应的教科书机制，但 mRNA 翻译和衰变的转录调控是硬币的另一面，对于快速转录至关重要。 和适应性刺激引起的反应。例如，免疫激活产生一个典型的时间 细胞因子 mRNA 的表达模式称为脉冲响​​应，这是一种快速的脉冲状增加和 表达量减少。细胞因子 mRNA 的不稳定是正确解决脉冲所必需的 反应和预防过量的细胞因子产生。人们越来越认识到RNA结合蛋白 （RBP）通过调节在实现适当的刺激诱导的基因表达反应中发挥关键作用 目标 RNA 的 RNA 加工、衰变和翻译。我们研究计划的总体目标是 了解 RBP 如何暂时协调刺激诱导的基因表达和细胞表型。 我们将使用该模型系统来研究动态 RBP-RNA 调节相互作用如何在时间上协调 刺激诱导的基因表达是人类肾上腺类固醇生成。这是一个经典的配体诱导系统 小肽血管紧张素 II (AngII) 与其在肾上腺肾小球带细胞中的受体结合，以刺激 醛固酮的产生，醛固酮是血压的主要调节剂。因此，严格控制监管时机 这是必需的，因为它必须响应 AngII 而从胆固醇快速产生。我们最近有 证明 AngII 对永生化和原代肾上腺皮质细胞的治疗会导致冲动 回复。此外，我们发现特定的 RBP 和受调控的 RNA 衰减有助于快速实施 和 AngII 脉冲响应的分辨率。在本提案中，我们将测试 MSI2 和 ZFP36L2 的假设 是分别增强和减弱刺激诱导的 mRNA 动力学的反作用力 通过调节目标 mRNA 的衰减和翻译水平，以确保正确的时间和幅度 脉冲响应。我们将利用尖端方法来获得时间和定量的见解 刺激诱导的 RBP 结合变化决定了靶标 mRNA 表达反应的变化， 最终细胞表型。我们使用的创新方法将有助于将该领域从静态转变为动态 RBP-mRNA 相互作用的动态视图，并获得有关 RBP-mRNA 如何调节的机制见解 相互作用控制 mRNA 命运决定，从而控制细胞反应。