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.

项目摘要 细胞适应多种压力条件的能力在各种生理和 病理环境，包括发育，癌症和神经系统疾病。我们的小组报告了 鉴定应力诱导的低复杂性二核苷酸重复反复非编码RNA，这些RNA衍生自刺激特异性 核糖体间间隔物（RigsRNA）的基因座；历史上人类基因组的神秘地区 被视为“垃圾” DNA。我们表明，低复杂度rigsRNA启动物理淀粉样蛋白生成 将核仁转化为淀粉样物体的程序：可逆的纤维膜细胞器 具有淀粉样蛋白样特征的固定蛋白质。虽然已经描述了许多无膜隔室 作为液体样（例如，应力颗粒，p-bodies，生殖细胞颗粒），发现了淀粉样体的发现 淀粉样生成过程的证据可以将生物学物理过渡到固体状态。 这种不寻常的翻译后调节途径使一系列阵列的迅速存储 淀粉样体中的内源性蛋白质抑制对严重环境反应的细胞中的代谢 侮辱。我们建议将2015年资助的NIGMS R01赠款转换为2018年的MIRA 统一主题“在压力期间低复杂性rigsRNA的功能”。我们的研究计划包括深入 了解（i）RIGSRNA激活生理淀粉样生成的机制 淀粉样物体和（ii）在应力期间rigsRNA和淀粉样体的功能。概述了这一点的研究 建议将涉及孤立的淀粉样体，主动翻译位点的多色单分子成像 在压力，蛋白质动力学，未拟合RNA，rRNA生物学和体外纤颤的长期读取测序 了解低复杂性RNA的细胞和生化功能在厌氧中的细胞中的细胞和生化功能 新陈代谢，以及其他条件。这项由NIGMS资助的研究使我们的实验室能够使 概念上的进步在我们对基因组中简单二核苷酸低复杂性重复的理解中取得了进步。第一的， Rigrsna构建淀粉样体的发现提供了证据，表明细胞可以激活生理 液体到固相的转变，以淀粉样蛋白样性质组装冷凝物。这些特征 来自众多的液体冷凝物的不同淀粉样物体，这些液体冷凝物填充了哺乳动物细胞， 通常不显示淀粉样蛋白生成特征。我们建议的工作不仅会阐明自适应 压力源的机制，但也为病理淀粉样生成的研究提供了替代见解 参与一系列人类神经系统疾病。其次，发现低复杂性RNA序列的发现 是RIGSRNA的功能决定剂可能会刺激对长二核苷酸的身体作用的研究 在整个基因组中观察到的基因间重复，但通常被视为无用的DNA/RNA。因此，这个 对对压力源的细胞反应感兴趣的科学家，项目将具有一般性的兴趣，长期不编码RNA 生物学，核/细胞质结构，翻译和物理/病理淀粉样蛋白发生。