Translation control of stress response and innate immunity

应激反应和先天免疫的翻译控制

• 批准号: 10004111
• 负责人: HYUNG D RYOO
• 金额: $ 34.08万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-20 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10004111
• 关键词: 5' Untranslated Regions; ATF6 gene; Affect; Animal Model; Animals; Attenuated; Bacterial Infections; Cells; Cellular Stress; Cellular Stress Response; Development; Diabetes Mellitus; Disease; Drosophila genus; EIF4EBP1 gene; Endoplasmic Reticulum; Gene Expression; Gene Expression Regulation; Genetic Transcription; Genetic Translation; Goals; Health; Human; Impairment; Inflammation; Innate Immune Response; Internal Ribosome Entry Site; Longevity; Mammals; Mediating; Messenger RNA; Metabolic Diseases; Molecular Genetics; Natural Immunity; Neurodegenerative Disorders; Nutrient; Open Reading Frames; Pathogenicity; Pathologic; Pathway interactions; Peptide Initiation Factors; Phenotype; Phosphorylation; Phosphotransferases; Physiological; RNA interference screen; Regulation; Resistance; Stress; Stress Response Signaling; Structure; Structure of beta Cell of islet; Study models; Testing; Therapeutic; Time; Transcript; Translational Regulation; Translational Repression; Translations; antimicrobial peptide; attenuation; base; biological adaptation to stress; endoplasmic reticulum stress; experimental study; genomic tools; inhibitor/antagonist; insight; interest; mutant; novel; prevent; programs; response; ribosome profiling; transcription factor

Project Summary The long-term goal of this project is to understand how mRNA translational regulatory mechanisms control cellular stress response under physiological and pathological conditions. The specific pathway of interest is the Integrated Stress Response (ISR), which involves stress-responsive kinases that phosphorylate the α subunit of eIF2 to attenuate general translational initiation. More recently, we discovered that ISR inhibits mRNA translation through the induction of yet another translational inhibitor 4E-BP. At the same time, such conditions paradoxically stimulate the translation of transcription factors such as ATF4, due to the presence of regulatory upstream Open Reading Frames (uORFs) that precede the main ATF4 ORF. How stress responsive transcripts evade translational inhibition, or even undergo more active expression, in stressed cells is one of the major conceptual questions that remains poorly understood. Our preliminary studies using Drosophila have led us to a number of new insights to this question: These include our findings that (1) ISR boosts innate immune response to bacterial infection and anti-microbial peptides have 5’UTRs that can evade translational inhibition imposed by ISR signaling, (2) that previously unexpected factors regulate the activation of ISR, and (3) that ATF6 also has a 5’UTR that stimulates the main ORF translation in response to stress. Here I propose to use Drosophila to investigate the underlying regulatory mechanisms and determine how the newly identified ISR regulatory factors affect innate immune response, inflammation, and lifespan of Drosophila. We will supplement the molecular genetics-based approach with genomic tools such as ribosome profiling and structure-based modeling studies. Three Specific Aims will be pursued: (1) We will determine how ISR signaling enhances innate immune response, mainly focusing on the idea that anti-microbial peptide transcripts undergo mRNA translation through an unconventional mechanism to evade translational inhibition associated with ISR signaling. (2) Through the characterization of the novel ISR regulators that we have identified, we plan to determine how uORF containing transcripts increase their translation when general mRNA translation is suppressed, and examine how they affect phenotypes associated with abnormal ISR regulation. (3) We will test the hypothesis that ATF6 is another transcription factor that is regulated at the level of mRNA translation during ISR to mediate its transcriptional response. Notably, impairment or excessive stimulation of this pathway underlies various neurodegenerative and metabolic disorders in humans. Therefore, a better understanding of the regulatory mechanisms will not only advance our conceptual understanding of gene expression regulation in cells under stress but also may prompt the development of new strategies to modulate ISR signaling for therapeutic purposes.

项目摘要 该项目的长期目标是了解mRNA翻译调控机制如何控制 生理和病理条件下的细胞应激反应。感兴趣的具体途径是 整合应激反应（ISR），涉及应激反应激酶，其磷酸化 eIF 2减弱一般翻译起始。最近，我们发现ISR抑制mRNA翻译， 通过诱导另一种翻译抑制剂4 E-BP。与此同时，这样的条件 相反地，由于调节因子的存在， 上游开放阅读框（uORF），其位于主ATF 4 ORF之前。压力反应转录本 逃避翻译抑制，甚至经历更活跃的表达，在应激细胞中是一个主要的 一些概念性的问题仍然不为人所知。我们对果蝇的初步研究使我们得出了 对这个问题有许多新的见解：这些包括我们的发现：（1）ISR增强先天免疫反应 抗微生物肽具有5 'UTR，其可以逃避由细菌感染引起的翻译抑制， ISR信号转导，（2）以前未预料到的因素调节ISR的激活，（3）ATF 6也具有 5 'UTR，其刺激响应于应激的主要ORF翻译。在这里，我建议用果蝇来 研究潜在的调节机制，并确定新发现的ISR调节因子 影响果蝇的先天免疫反应、炎症和寿命。我们将补充分子 基于遗传学的方法与基因组工具，如核糖体分析和基于结构的建模研究。 本论文的主要目的是：（1）研究ISR信号通路如何增强先天免疫 反应，主要集中在这样的想法，即抗微生物肽转录物通过 一种非常规的机制，以逃避与ISR信号相关的翻译抑制。(2)通过 我们已经确定了新的ISR调节器的表征，我们计划确定如何uORF含有 当一般的mRNA翻译被抑制时，转录本增加了它们的翻译，并研究了它们是如何被抑制的。 影响与异常ISR调节相关的表型。(3)我们将测试ATF 6是另一种 在ISR期间在mRNA翻译水平上调节以介导其转录的转录因子 反应值得注意的是，该通路的损伤或过度刺激是各种神经退行性疾病的基础。 和人体代谢紊乱。因此，更好地了解监管机制不仅 推进我们对压力下细胞基因表达调节的概念性理解，但也可能促进 开发新的策略来调节ISR信号以用于治疗目的。