Investigating the dynamics of the neuronal RNP network

RNA结合蛋白(RBP)被认为是RNAs的主要调节者，通过组装成核糖核蛋白颗粒(RNPs)来控制其细胞命运。因此，限制性商业惯例实际上调节了细胞内RNA的所有步骤，从转录、剪接、修饰、细胞内定位到腐烂。然而，我们才刚刚开始了解RNP的组装以及这种组装如何在细胞内受到调控。在这项提议中，我们想要关注初级神经元中的限制性商业惯例网络。从双链RBP Staufen2作为RNA颗粒的关键成分开始，我们想研究RNAs是如何包装成RNP的。特别是，我们想着重于由于失去Staufen2而导致的RNP组成中的直接影响以及补偿影响。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Staufen2的已知相互作用因子。在这里，我们将利用一系列互动屏幕，并辅之以活细胞成像和最先进的生物化学，来揭示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RNPs的生命周期以及RBP网络如何对以RNA颗粒成分故障为特征的严重人类神经退行性疾病、神经和神经精神疾病的临床相关功能丧失表型做出反应的重要新见解。

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.