Exploring chaperone code control of TDP-43 function in ALS

探索 ALS 中 TDP-43 功能的伴侣代码控制

• 批准号: 10724923
• 负责人: Andrew William Truman
• 金额: $ 41.38万
• 依托单位: UNIVERSITY OF NORTH CAROLINA CHARLOTTE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-15 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10724923
• 关键词: Amyotrophic Lateral Sclerosis; Binding; Binding Proteins; Biological Process; Biology; Cells; Code; Collection; Communities; Complex; DNA-Binding Proteins; Data; Mammalian Cell; Mass Spectrum Analysis; Modification; Molecular Chaperones; Mutation; Nature; Neurodegenerative Disorders; Phosphorylation; Phosphorylation Site; Post-Translational Protein Processing; Property; Protein C; Protein Family; Protein Isoforms; Proteins; Proteomics; Regulation; Research; Role; Site; Specificity; Structure; Surface; TDP-43 aggregation; Therapeutic; Toxic effect; Work; Yeasts; amyotrophic lateral sclerosis therapy; crosslink; frontotemporal lobar dementia amyotrophic lateral sclerosis; novel; novel therapeutics; prevent; protein TDP-43; protein aggregation; protein folding; response; tool

Aggregation of the TAR DNA-binding protein (TDP-43) is associated with neurodegenerative disorders such as frontotemporal degeneration (FTD) and amyotrophic lateral sclerosis (ALS). Any strategy that alters TDP-43 aggregation, function and cellular toxicity may form the basis of potent novel ALS therapies. A major regulator of protein folding and aggregation in cells is the Hsp70 molecular chaperone. Research has primarily focused on how Hsp70 function specificity arises through regulation of a) expression of Hsp70, b) isoform differences in the Hsp70 protein family and c) the variety of co-chaperone proteins that bind to the Hsp70 molecule. Despite the proteomic identification of over seventy phosphorylation sites on both yeast and mammalian Hsp70, the biological function of most of these sites remains unknown. In this proposal, we intend to investigate the connection between post-translational modification (PTM) of Hsp70 and TDP-43 function. In particular, we will determine how chaperone PTMs are altered in response to the presence of TDP-43 in mammalian and yeast cells and how the modification of these PTMs impacts TDP-43 toxicity. Finally, we intend to use cross-linking mass spectrometry to analyze the interaction between Hsp70 and TDP-43 with a focus on the interaction surface between these two proteins. This information will provide ket preliminary data for larger studies that determine the mechanisms regulating these PTMs and their specific effect on TDP-43.

TAR DNA 结合蛋白 (TDP-43) 的聚集与神经退行性疾病相关，例如 如额颞叶变性（FTD）和肌萎缩侧索硬化症（ALS）。任何改变 TDP-43 的策略 聚集、功能和细胞毒性可能构成有效的新型 ALS 疗法的基础。主要监管者 Hsp70 分子伴侣是细胞内蛋白质折叠和聚集的重要组成部分。研究主要集中于 关于 Hsp70 功能特异性如何通过调节 a) Hsp70 的表达，b) 亚型差异而产生 Hsp70 蛋白家族和 c) 与 Hsp70 分子结合的各种辅助伴侣蛋白。尽管 对酵母和哺乳动物 Hsp70 上 70 多个磷酸化位点进行蛋白质组学鉴定， 大多数这些位点的生物学功能仍然未知。 在本提案中，我们打算研究翻译后修饰（PTM）之间的联系 Hsp70 和 TDP-43 功能。特别是，我们将确定如何改变伴侣 PTM 以响应 TDP-43 在哺乳动物和酵母细胞中的存在以及这些 PTM 的修饰如何影响 TDP-43 毒性。最后，我们打算利用交联质谱分析Hsp70和Hsp70之间的相互作用。 TDP-43 重点关注这两种蛋白质之间的相互作用表面。此信息将提供 确定调节这些 PTM 的机制及其具体效果的大型研究的初步数据 在 TDP-43 上。