Investigating the dynamics of the neuronal RNP network

RNA结合蛋白（RBP）被认为是RNA的主要调节因子，通过组装成核糖核蛋白颗粒（RNP）来控制RNA的细胞命运。因此，RBP实际上调节细胞内RNA的所有步骤，从转录、剪接、修饰、细胞内定位直到衰变。然而，我们才刚刚开始了解RNP的组装以及它在细胞内是如何被调节的。在这个提议中，我们希望专注于初级神经元中的RBP网络。从双链RBP Staufen2作为关键的RNA颗粒组分开始，我们想研究RNA如何包装成RNP。特别是，我们想把重点放在直接以及RNP组合物的补偿效应所造成的损失施陶芬2。因此，将特别强调RNP网络内的其他RNP，例如Ago2、DDX6和Pumilio2，它们都是Staufen 2的已知相互作用物。在这里，我们将利用一组相互作用屏幕，辅以活细胞成像和最先进的生物化学，以揭示RBP网络如何动态组装和平衡分子和细胞的变化。总之，这无疑将导致对RNP生命周期的重要新见解，以及RBP网络如何对以RNA颗粒组分故障为标志的严重人类神经退行性、神经和神经精神疾病中的临床相关功能丧失表型作出反应。

RNA-binding proteins (RBPs) are thought to be major regulators of RNAs, controlling their cellular fate by assembly into ribonucleoproteinparticles (RNPs). RBPs thereby regulate virtually all steps of an RNA inside a cell, ranging from transcription, splicing, modification, intracellular localization, up to decay. Yet, we are only beginning to understand RNP assembly and how this might be regulated inside cells. In this proposal, we want to focus on the network of RBPs in primary neurons. Starting from the double-stranded RBP Staufen2 as a key RNA granule component, we would like to investigate how RNAs are packaged into RNPs. In particular, we would like to focus on direct as well as also compensatory effects in RNP composition caused by loss of Staufen2. A particular emphasis will thereby be given on other RNPs within the RNP network, e.g. Ago2, DDX6 and Pumilio2, all known interactors of Staufen2. Here, we will exploit a battery of interactor screens complemented with live-cell imaging and state-of-the-art biochemistry to unravel how the RBP network dynamically assembles and balances molecular and cellular alterations. Together, this will undoubtedly result in important new insight into the life cycle of RNPs and how the RBP network reacts to clinically relevant loss-of-function phenotypes in severe human neurodegenerative, neurological and neuropsychiatric diseases that are hallmarked by malfunctions of RNA granule components.