Elucidating the Mechanism in the Regulation of RNA-binding Protein Phase Separation

阐明 RNA 结合蛋白相分离的调节机制

• 批准号: 10028997
• 负责人: Lin Guo
• 金额: $ 39万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10028997
• 关键词: Biogenesis; Biological; Biological Process; Cells; Disease; Fluorescence; Foundations; Goals; Health; Knowledge; Link; Liquid substance; Molecular; Nerve Degeneration; Neurodegenerative Disorders; Nuclear Import; Nucleic Acids; Oligonucleotides; Organelles; Pathologic; Phase; Phase Transition; Proteins; RNA; RNA-Binding Proteins; Regulation; Research; Specificity; Therapeutic; Work; base; biophysical techniques; combat; driving force; innovation; novel therapeutics; programs; protein aggregation; proteotoxicity; receptor; single molecule

Abstract: Liquid–liquid phase separation of RNA-binding proteins (RBP) is a recently appreciated means of intracellular compartmentalization underpins the biogenesis of diverse membraneless organelles. Despite clear biological utility, dysregulated phase separation of RNA-binding protein (RBP) leads to protein aggregation and fibrils formation, which are key pathological features of numerous neurodegenerative diseases. The central hypothesis of my work, and the driving force of my lab, is that reversing aberrant phase transition and aggregation is a potential avenue to combat these fatal diseases. In fact, we and others have established that nuclear import receptors (NIRs) such as Kapβ2 can reverse protein aberrant phase transition and elevated Kapβ2 expression can suppress neurodegeneration caused by disease-linked RNA-binding proteins. Thus, NIRs and other regulators of protein phase transition could be leveraged therapeutically to restore normal function of membraneless organelles and mitigate proteotoxicity caused by aberrant phase transition. However, several key knowledge gaps regarding the regulation of phase transition must be filled, to lay the foundation to develop such therapeutic strategies. Thus the long-term goal of my research program is to achieve a more comprehensive understanding of the regulatory mechanism of protein phase separation and leverage our understanding of phase regulation to develop strategies with therapeutic potential to reverse proteotoxicity induced by aberrant phase transition. I seek to elucidate the molecular mechanism of the regulation of protein phase separation, both in health and disease conditions. We propose to focus on three main themes over the next five years: (1) Define the mechanism of how Kapβ2 reverses protein aberrant phase transition. (2) Define the scope and specificity of Kapβ2 and other nuclear import receptors in their function in regulating phase separation in live cells. (3) Based on our preliminary results that RNA oligonucleotides with specific sequences have diverse activities in regulating RNA-binding protein phase separation, we will identify the sequence space of RNA in regulating RBP phase separation and develop innovative single-molecule fluorescence-based biophysical methods to understand the mechanism of RNA’s function. Our work will significantly contribute to our understanding of the regulatory mechanism of phase separation in cells and how the breakdown of this regulation leads to disease conditions. Furthermore, this work will set the stage for developing strategies to enhance the activity of Kapβ2, as well as RNA-based oligonucleotides that can mitigate toxic aberrant phase transition, both of which provide the basis for developing innovative therapeutics to restore the healthy protein phase in cells.

抽象的： RNA 结合蛋白 (RBP) 的液-液相分离是最近受到重视的细胞内分离方法 尽管有明确的生物学意义，区室化还是多种无膜细胞器生物发生的基础。 RNA 结合蛋白 (RBP) 的实用性、失调的相分离导致蛋白质聚集和原纤维 形成，这是许多神经退行性疾病的关键病理特征。 我的工作的重点，以及我的实验室的驱动力，是逆转异常的相变和聚集是一个 事实上，我们和其他人已经确定了核进口。 Kapβ2 等受体 (NIR) 可以逆转蛋白质异常相变和 Kapβ2 表达升高 可以抑制由疾病相关的 RNA 结合蛋白引起的神经变性，例如 NIR 和其他蛋白。 蛋白质相变的调节剂可以在治疗上用来恢复正常功能 然而，有几个关键点。 必须填补有关相变调控的知识空白，为开发此类技术奠定基础 因此，我的研究计划的长期目标是实现更全面的治疗策略。 了解蛋白质相分离的调控机制并利用我们的理解 相位调节以开发具有治疗潜力的策略，以逆转异常引起的蛋白毒性 我试图阐明蛋白质相分离调节的分子机制。 在健康和疾病状况下。 我们建议未来五年重点关注三个主题：（1）定义 Kapβ2 的机制 逆转蛋白质异常相变 (2) 定义 Kapβ2 和其他核输入的范围和特异性。 (3) 根据我们的初步结果 具有特定序列的RNA寡核苷酸在调节RNA结合蛋白相方面具有多种活性 分离，我们将确定RNA在调节RBP相分离中的序列空间并开发 基于创新单分子荧光的生物物理方法来了解 RNA 的机制 我们的工作将极大地促进我们对相调控机制的理解。 此外，这项工作还研究了细胞内的细胞分离以及这种调节的破坏如何导致疾病。 将为制定增强 Kapβ2 以及基于 RNA 的活性的策略奠定基础 可以减轻有毒异常相变的寡核苷酸，两者都为开发提供了基础 恢复细胞健康蛋白质状态的创新疗法。