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 细胞适应各种应激条件的能力在各种生理和生理环境中起着至关重要的作用。 病理环境，包括发育、癌症和神经疾病。我们的小组报告了 应激诱导的刺激特异性低复杂性二核苷酸重复非编码RNA的鉴定 核糖体基因间隔区(RIGSRNA)；历史上人类基因组的一个谜团区域 被斥为“垃圾”DNA。我们发现低复杂性rIGSRNA启动生理性淀粉样变性 将核仁转化为淀粉样体的程序：由以下组成的可逆纤维状无膜细胞器 具有淀粉样蛋白特征的固定化蛋白。虽然已经描述了许多无膜隔室 作为液体样(如应激颗粒、P小体、生殖细胞颗粒)，淀粉样体的发现提供了 淀粉样变性过程的证据，可以从生理上将生物物质转变为类固体状态。 这一相当不寻常的翻译后调控途径使一系列 淀粉样体中的内源性蛋白抑制细胞对恶劣环境的反应 侮辱。我们建议将2015年资助并于2018年续期的NIGMS R01赠款转换为MIRA 统一主题“低复杂性rIGSRNA在应激中的作用”。我们的研究项目包括深入的 研究了解(I)rIGSRNA激活生理淀粉样蛋白形成的机制 淀粉样体；(Ii)rIGSRNA和淀粉样体在胁迫中的作用。这篇文章中概述的研究 该提案将涉及分离的淀粉样体，活性翻译位点的多颜色单分子成像 应激、蛋白质动力学、非模板rna的长阅读测序、rRNA生物学和体外纤颤 了解低复杂性RNA在厌氧细胞中的细胞和生化功能的方法 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 在我们理解简单二核苷酸低复杂性在基因组中重复的概念上的进展。第一, RIGRSNA构建淀粉样体的发现为细胞可以激活生理活性提供了证据 液体到固体的相变，以组装具有淀粉样属性的冷凝物。这些特点 将淀粉样体与大量存在于哺乳动物细胞中的液体凝结物区分开来，这些凝集物 通常不显示淀粉样变性特征。我们提议的工作不仅将阐明自适应 应激源的机制，也为病理性淀粉样变的研究提供了新的见解 参与了一系列人类神经疾病。第二，发现低复杂性的RNA序列 RIGSRNA的功能决定因素可能会刺激对长二核苷酸生理作用的研究 在基因组中观察到的基因间重复，但通常被认为是无用的DNA/RNA。因此，这是 该项目将引起对细胞对应激源的反应感兴趣的科学家的普遍兴趣，应激源是长的非编码RNA 生物学，核/细胞质结构，翻译和生理性/病理性淀粉样变。