Regulation of Alternative Splicing in the Brain by a Large Assembly of Splicing Regulators

项目总结/摘要 选择性剪接是塑造大脑功能复杂性的关键过程。因此，许多 神经和精神疾病是由RNA结合蛋白或其靶点突变引起的， 选择性剪接RNA结合蛋白Rbfox家族在神经元细胞中调节选择性剪接 发育及其突变与自闭症谱系障碍和各种家族性疾病有关。 癫痫病为了了解Rbfox在这些疾病中的作用，许多研究都集中在如何 单个Rbfox蛋白质与它们的靶结合并调节它们的剪接。然而，Rbfox蛋白也是 已知与影响其功能的辅因子相互作用。我们以前发现，核中的Rbfox蛋白， 小鼠脑几乎专门结合于大的剪接调节子组装体（LASR）。但 神经元LASR复合物的完整亚基是什么以及它们如何与Rbfox相互作用， 调节神经元中的选择性剪接。在本提案中，我将定义LASR的神经元成分， 确定小鼠脑中LASR/Rbfox复合物及其亚基的转录组范围的靶点，以及 分析LASR的组分如何影响Rbfox在神经元中的剪接调节活性。这些研究将 阐明在一种新的剪接调节蛋白中RNA结合蛋白之间的组合相互作用 复杂的形状的基因调控电路的大脑，并将进一步我们了解的功能， 与癫痫和自闭症谱系障碍有关的Rbfox蛋白质。

PROJECT SUMMARY/ABSTRACT Alternative splicing is a key process in shaping the functional complexity of the brain. Accordingly, many neurologic and psychiatric disorders are caused by mutations in RNA binding proteins or their targets in alternative splicing. The Rbfox family of RNA Binding proteins regulate alternative splicing during neuronal development and their mutations have been implicated in autism spectrum disorder and various familial epileptic disorders. To understand the role of Rbfox in these diseases, many studies have focused on how individual Rbfox proteins bind to their targets and regulate their splicing. However, Rbfox proteins are also known to interact with cofactors that influence their function. We previously found that nuclear Rbfox proteins in the mouse brain are almost exclusively bound to a large assembly of splicing regulators (LASR). However, it remains unknown what the full subunits of the neuronal LASR complex are and how they interact with Rbfox to regulate alternative splicing in neurons. In this proposal, I will define the neuronal components of LASR, determine the transcriptome-wide targets of the LASR/Rbfox complex and its subunits in the mouse brain, and analyze how components of LASR affect Rbfox’s splicing regulatory activity in neurons. These studies will elucidate how combinatorial interactions between RNA binding proteins in a novel splicing regulatory protein complex shape the gene regulatory circuit of the brain and will further our understanding of the function of the Rbfox proteins which have been implicated in epilepsy and autism spectrum disorder.