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，也是与年龄相关的神经变性的标志，包括阿尔茨海默病和帕金森病。处理聚集体的基本机制主要依赖于蛋白酶体介导的降解和选择性自噬途径。相反，细胞对极端蛋白质毒性应激的反应是积极地将聚集体运送到微管组织中心，在那里它们结合成一个专门的细胞器，称为聚集体。该过程依赖于负端定向电机动力。通过这项研究，我将揭示将动力蛋白招募到聚集体并将其靶向聚集体所需的因素网络，并研究马达是否也在聚集的基础清除中发挥作用。在绝大多数情况下，动力蛋白转运是由专门的动力蛋白激活适配器介导的。我将使用质谱法和计算方法来确定哪个激活适配器介导动力蛋白聚集，并使用生物化学和细胞生物学验证相互作用。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聚集，并确认它们的功能与消耗。我将调查在基础条件下运输途径是否仍然活跃。我将结合使用AlphaFold和Cryo-EM，对连接聚合体和细胞马达的连接提供全面的分子理解。这项研究的成果将对研究蛋白质停滞、神经变性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.