DynProtect - Mechanisms of dynein-dependent transport and degradation of protein aggregates.

蛋白质聚集体的积累是对真核细胞的主要威胁，也是老年性神经变性的标志，包括阿尔茨海默氏症和帕金森氏病。处理聚集体的基本机制很大程度上取决于蛋白酶体介导的降解和选择性自噬的途径。相比之下，细胞对极端的蛋白毒性压力的反应是积极地将聚集体运输到微管组织中心，在那里它们聚集成一个特殊的细胞器，称为侵袭体。这一过程依赖于负端定向运动动力蛋白。有了这份奖学金，我将揭示招募动力蛋白到聚集体并将它们瞄准攻击者所需的因素网络，并解决运动是否也在聚集的基本清除中发挥作用。在绝大多数情况下，动力蛋白的运输是由专门的动力蛋白激活适配器介导的。我将使用质谱学和计算方法来确定哪个激活接头介导动力蛋白重新聚集到聚集体中，并使用生物化学和细胞生物学来验证这种相互作用。在神经元中使用活细胞成像来识别哪些蛋白质在运输过程中被招募到聚集体中，并确认它们在耗尽时的功能。我将调查在基本条件下运输路径是否仍然活跃。我将使用AlphaFold和Cryo-EM的组合来提供对连接聚集体和细胞马达的连接的全面分子理解。这项研究的成果将对研究蛋白质稳定性、神经退行性变和细胞内运输的社区产生极大的兴趣。

Accumulation of protein aggregates is a major threat to eukaryotic cells, and a hallmark of age-associated neurodegeneration, including Alzheimer's and Parkinson's disease. Basal mechanisms to dispose of aggregates largely rest on the pathways of proteasome-mediated degradation and selective autophagy. In contrast, cells react to extreme proteotoxic stress by actively transporting aggregates to the microtubule organising centre, where they coalesce into a specialised organelle, named the aggresome. The process depends on the minus-end directed motor dynein. With this fellowship, I will uncover the network of factors required to recruit dynein to aggregates and target them to the aggresome, and address whether the motor also plays a role in the basal clearance of aggregation.Dynein transport is in the vast majority of cases mediated by specialised dynein activating adaptors. I will use mass spectrometry and computational methods to identify which activating adaptor mediates dynein recruitment to aggregates, and validate the interaction using biochemistry and cell biology. use live-cell imaging in neurons to identify which proteins are recruited to aggregates during transport and confirm their function with depletions. I will investigate whether transport pathways are still active under basal conditions. I will use a combination of AlphaFold and Cryo-EM to provide a comprehensive molecular understanding of the connections that link aggregates to cellular motors. The outputs of this fellowship will be of great interest to communities researching proteostasis, neurodegeneration and intracellular transport.