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.

摘要 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 决议。尽管转录反应是一种教科书上的刺激诱导反应机制，但后 转录调控的mRNA翻译和衰退是硬币的另一面，是必不可少的快速 以及适应性刺激诱导的反应。例如，免疫激活会产生一个典型的时间 细胞因子mRNAs的表达模式称为脉冲反应，这是一种快速的脉冲状增加和 表达减少。细胞因子mRNAs的失稳是正确解决冲动所必需的。 细胞因子过度产生的反应和预防。人们越来越认识到，RNA结合蛋白 限制性商业惯例(RBPs)在实现适当的刺激诱导的基因表达反应中发挥着关键作用 靶RNA的加工、衰变和翻译。我们研究计划的总体目标是 了解限制性商业惯例如何在时间上协调刺激诱导的基因表达和细胞表型。 我们将用来研究动态RBP-RNA调控相互作用如何在时间上协调的模型系统 刺激诱导的基因表达是人类肾上腺类固醇激素的合成。这是一个经典的配基诱导系统 小肽血管紧张素II(AngII)与肾上腺肾小球带细胞上的受体结合刺激 生产血压的主要调节剂--醛固酮。因此，严格控制监管时机是 这是必需的，因为它必须由胆固醇迅速从头产生，以应对血管紧张素Ⅱ。我们最近做了 证明血管紧张素转换酶治疗永生化和原代肾上腺皮质细胞可产生一种冲动 回应。此外，我们发现了特定的限制性商业惯例和受调控的RNA衰变有助于快速实施 以及血管紧张素转换酶脉冲反应的分辨率。在这个提议中，我们将检验MSI2和Zfp36l2的假设 是分别增强和减弱刺激诱导的信使核糖核酸的动力学的反作用力 通过调节靶mRNA的衰变和翻译来确保正确的时机和幅度 冲动反应。我们将利用尖端方法来获得时间和数量上的洞察 刺激诱导的RBP结合变化决定了靶mRNA表达反应的变化和 最终是细胞表型。我们使用的创新方法的使用将有助于将该领域从静态转向 动态观察RBP-信使核糖核酸的相互作用，并实现对RBP-信使核糖核酸调控机制的洞察 相互作用控制着信使核糖核酸的命运决定，从而控制细胞的反应。