Exploring chaperone code control of TDP-43 function in ALS

TAR DNA结合蛋白（TDP-43）的聚集与神经退行性疾病有关， 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形成有效的新型ALS疗法的基础。一个主要的监管机构 Hsp 70分子伴侣是细胞内蛋白质折叠和聚集的关键。研究主要集中在 关于Hsp 70功能特异性如何通过调节a）Hsp 70的表达，B）Hsp 70的同种型差异， Hsp 70蛋白家族和c）结合Hsp 70分子的多种共伴侣蛋白。尽管 酵母和哺乳动物Hsp 70上超过70个磷酸化位点的蛋白质组学鉴定， 大多数这些位点的生物学功能仍然未知。 在这个建议中，我们打算调查翻译后修饰（PTM）之间的联系， Hsp 70和TDP-43起作用。特别地，我们将确定伴侣蛋白PTM如何响应于蛋白质的改变而改变。 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，我们打算使用交联质谱分析热休克蛋白70和 TDP-43的重点是这两种蛋白质之间的相互作用表面。这些信息将为Ket 确定这些PTM调控机制及其具体作用的大型研究的初步数据 在TDP-43上

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.