Deciphering the roles of neurite-targeted RBPs in mRNA localization and local translation

RNA定位和翻译是体轴建立、细胞迁移和突触可塑性的重要生物学过程。它们由RNA结合蛋白（RBP）介导，所述RNA结合蛋白与mRNA中的特异性顺式调节元件结合并介导mRNA的定位、翻译和稳定性。然而，到目前为止，很少有全面的研究本地RBP及其靶基因在哺乳动物细胞。利用神经突/索马分离技术，我们对小鼠胚胎干细胞分化为神经元的局部蛋白质组和转录组进行了全基因组筛选。该应用程序是在SPP 1935（解密mRNP代码：转录后基因调控的RNA结合决定因素）的框架内，因此在这里，我们专注于我们筛选中鉴定的富含神经突的RBP（mRNA相互作用组）。我们建议解剖这些RBPs的作用，在本地化，本地翻译和稳定性的目标，使用CLIP和RIP分析，结合空间蛋白质组学，转录组学和核糖体分析的神经突和索马车厢。这项研究有可能大大提高我们对mRNP在空间维度上功能的理解。

RNA localization and local translation are important biological processes that underlie establishment of body axis, cell migration and synaptic plasticity. They are mediated by RNA binding proteins (RBPs) binding to specific cis-regulatory elements in mRNAs and mediating their localization, translation and stability. However, so far there have been few comprehensive studies of localized RBPs and their target mRNAs in mammalian cells. Using neurite/soma fractionation scheme, we have conducted a genome-wide screen for local proteome and transcriptome of neurons differentiated from mouse embryonic stem cells. This application is within the frame of SPP 1935 (Deciphering the mRNP code: RNA-bound Determinants of Post-transcriptional Gene Regulation), therefore here we are focusing on neurite-enriched RBPs (mRNA-interactome group) identified in our screen. We propose to dissect the roles of these RBPs in localization, local translation and stability of their targets, using CLIP and RIP assays, combined with spatial proteomic, transcriptomic and ribosome profiling analyses of neurite and soma compartments. The proposed research has the potential to substantially advance our understanding of mRNP function in spatial dimension.