The Nucleolar Detention Center: A Hub of Long Noncoding RNA that Imprison Proteins during Stress

核仁拘留中心：在压力下囚禁蛋白质的长非编码 RNA 中心

• 批准号: 10622035
• 负责人: Stephen Lee
• 金额: $ 38.38万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10622035
• 关键词: Amyloid; Biochemical; Biological; Biological Assay; Biology; Cells; Characteristics; Color; Cytoplasmic Structures; DNA; Development; Dinucleoside Phosphates; Dinucleotide Repeats; Disease; Funding; Genome; Germ Cells; Grant; Human; Human Genome; Immobilization; Imprisonment; In Vitro; Junk DNA; Laboratories; Liquid substance; Malignant Neoplasms; Mammalian Cell; Membrane; Metabolism; National Institute of General Medical Sciences; Nuclear; Organelles; Pathologic; Pathway interactions; Phase Transition; Physical condensation; Physiological; Play; Process; Property; Protein Dynamics; Proteins; RNA; RNA Sequences; Regulatory Pathway; Reporting; Research; Ribosomal RNA; Ribosomes; Role; Scientist; Site; Solid; Stimulus; Stress; Translations; Untranslated RNA; Work; amyloidogenesis; detention center; granule cell; insight; interest; molecular imaging; nervous system disorder; programs; response; single molecule; stress granule; stressor

Project Summary The ability of cells to adapt to a wide variety of stress conditions plays a critical role in various physiological and pathological settings, including development, cancer and neurological disorders. Our group reported the identification of stress-induced low complexity dinucleotide repeat noncoding RNA derived from stimuli-specific loci of the ribosomal intergenic spacer (rIGSRNA); an enigmatic region of the human genome historically dismissed as “junk” DNA. We showed that low complexity rIGSRNA initiate physiological amyloidogenic programs that convert nucleoli into Amyloid-bodies: reversible fibrous membrane-less organelles composed of immobilized proteins with amyloid-like features. While many membrane-less compartments have been described as liquid-like (e.g., stress granules, P-bodies, germ cell granules), the discovery of Amyloid-bodies provided evidence of an amyloidogenic process that can physiologically transition biological matter to a solid-like state. This rather unusual post-translational regulatory pathway enables the rapid and reversible storage of an array of endogenous proteins in Amyloid-bodies to suppress metabolism in cells responding to severe environmental insults. We propose to convert our NIGMS R01 grant funded in 2015 and renewed in 2018 to a MIRA under the unifying theme “Function of low complexity rIGSRNA during stress”. Our research program includes in-depth studies to understand (i) the mechanisms by which rIGSRNA activate physiological amyloidogenesis to construct Amyloid-bodies and (ii) the function of rIGSRNA and Amyloid-bodies during stress. Studies outlined in this proposal will involve isolated Amyloid-bodies, multi-color single molecule imaging of active translation sites during stress, protein dynamics, long read sequencing of untemplated RNA, rRNA biology, and in vitro fibrillation assays to understand the cellular and biochemical functions of low complexity RNA in cells engaging in anaerobic metabolism, amongst other conditions. This NIGMS-funded research has enabled our laboratory to make conceptual advances in our understanding of simple dinucleotide low complexity repeats in the genome. First, the discovery that rIGRSNA construct Amyloid-bodies provided evidence that cells can activate physiological liquid-to-solid phase transitions to assemble condensates with amyloid-like properties. These characteristics distinguish Amyloid-bodies from the multitude of liquid condensates that populate mammalian cells, which typically do not display amyloidogenic features. Our proposed work will not only shed light on adaptive mechanisms to stressors but also provide alternative insights for the study of pathological amyloidogenesis involved in an array of human neurological disorders. Second, the finding that low complexity RNA sequences are functional determinants of rIGSRNA may stimulate research on the physiological role of long dinucleotide intergenic repeats observed across the genome, but generally dismissed as useless DNA/RNA. Hence, this project will be of general interest to scientists interested in cellular response to stressors, long noncoding RNA biology, nuclear/cytoplasmic structures, translation and physiological/pathological amyloidogenesis.

项目概要 细胞适应多种应激条件的能力在各种生理和心理过程中起着至关重要的作用。 我们的小组报告了病理情况，包括发育、癌症和神经系统疾病。 鉴定源自刺激特异性的应激诱导的低复杂性二核苷酸重复非编码 RNA 核糖体基因间隔区（rIGSRNA）的位点；人类基因组历史上的一个神秘区域； 我们发现低复杂性的 rIGSRNA 会引发生理性淀粉样蛋白生成。 将核仁转化为淀粉样蛋白体的程序：由以下组成的可逆纤维膜无细胞器 虽然已经描述了许多无膜隔室，但具有淀粉样蛋白样特征的固定化蛋白质。 作为液体状（例如应激颗粒、P 体、生殖细胞颗粒），淀粉样蛋白体的发现提供了 淀粉样蛋白生成过程的证据，该过程可以在生理上将生物物质转变为固体状态。 这种相当不寻常的翻译后调节途径能够快速且可逆地存储一系列 淀粉样蛋白体中的内源蛋白可抑制细胞响应恶劣环境的代谢 我们建议将 2015 年资助并于 2018 年更新的 NIGMS R01 赠款转换为 MIRA。 我们的研究计划包括深入的统一主题“低复杂性 rIGSRNA 在压力下的功能”。 研究旨在了解 (i) rIGSRNA 激活生理性淀粉样变形成的机制 淀粉样蛋白体以及 (ii) rIGSRNA 和淀粉样蛋白体在应激过程中的功能研究概述。 提案将涉及分离的淀粉样蛋白体、活性翻译位点的多色单分子成像 应激期间、蛋白质动力学、非模板 RNA 的长读测序、rRNA 生物学和体外纤维颤动 了解厌氧细胞中低复杂性 RNA 的细胞和生化功能的测定 这项由 NIGMS 资助的研究使我们的实验室能够实现新陈代谢等条件。 我们对基因组中简单二核苷酸低复杂性重复的理解在概念上取得了进展。 rIGRSNA 构建淀粉样蛋白体的发现提供了细胞可以激活生理学的证据 液-固相变以组装具有类淀粉样特性的冷凝物。 将淀粉样蛋白体与哺乳动物细胞中的大量液体冷凝物区分开来， 通常不表现出淀粉样蛋白形成特征，我们提出的工作不仅会揭示适应性。 应激源机制，同时也为病理性淀粉样蛋白生成的研究提供替代见解 第二，低复杂性RNA序列的发现。 rIGSRNA 的功能决定因素可能会促进对长二核苷酸生理作用的研究 在整个基因组中观察到基因间重复，但通常被认为是无用的 DNA/RNA。 该项目将引起对细胞对应激源、长非编码RNA的反应感兴趣的科学家的普遍兴趣 生物学、核/细胞质结构、翻译和生理/病理淀粉样蛋白生成。