Molecular characterization of a novel function of the RNA-binding protein Rbfox

描述(由申请人提供)：转录后基因调节已经成为一种关键的控制机制，在发育和成年期协调不同的细胞过程。RNA结合蛋白在RNA代谢中起着不可或缺的作用，调节包括剪接和翻译在内的一系列过程。其中一个家族是RNA结合狐狸(RBFox)蛋白，其成员通过高度保守的RNA识别基序与(U)GCAUG元件相互作用。特别是，RBFOX1与从神经系统疾病到癌症的许多人类疾病有关。RBFOX1基因编码几种异构体：调节组织特异性选择性剪接的核异构体和细胞质异构体，其功能尚不清楚。在果蝇种系中，RBFOX1存在于细胞质中，而专为减少细胞质RBFOX1而设计的RNAi会导致种系分化受损。我们已经证明RBFOX1抑制了3‘UTRs中含有(U)GCAUG元件的转录本的翻译。令人兴奋的是，我们的结果为广泛研究的剪接调控因子提供了一种新的功能，并为这一提议提供了基础。这个 这项建议的主要目的是加深对RBFOX1的分子和发育功能的理解。我们的总体策略是使用果蝇特有的复杂遗传和分子工具来研究RBFOX1在体内的功能。我们将利用我们分离的RBFOX1突变株从遗传学上研究RBFOX1控制翻译的分子机制。最后，我们将探索细胞质RBFOX1负性调节生殖系内钙反应的可能性。我们预计，这些努力将提高我们对人类RBFOX1功能的了解，并可能提高其在人类发病机制中的作用。

DESCRIPTION (provided by applicant): Post-transcriptional gene regulation has emerged as a critical control mechanism that orchestrates diverse cellular processes during development and adulthood. RNA binding proteins play integral roles in RNA metabolism, regulating a range of processes including splicing and translation. One such family is the RNA-binding fox (Rbfox) proteins, members of which interact with (U)GCAUG elements through a highly conserved RNA-recognition motif. In particular, RBFOX1 has been linked with a number of human disorders ranging from neurological diseases to cancer. The RBFOX1 gene encodes several isoforms: nuclear isoforms that regulate tissue specific alternative splicing and cytoplasmic isoforms, the function of which remains unclear. In the Drosophila germline, Rbfox1 is observed in the cytoplasm and RNAi designed to specifically reduce cytoplasmic Rbfox1 results in impaired germline differentiation. We have shown that Rbfox1 represses translation of transcripts containing (U)GCAUG elements within their 3' UTRs. Excitingly, our results suggest a novel function for an extensively studied splicing regulator and provide the basis for this proposal. The main goal of this proposal is to provide a deeper understanding of the molecular and developmental function of RBFOX1. Our general strategy is to use the sophisticated genetic and molecular tools unique to Drosophila to study Rbfox1 function in vivo. We will take advantage of a Rbfox1 mutant line that we have isolated to genetically investigate the molecular mechanism by which Rbfox1 controls translation. Lastly, we will explore the possibility that cytoplasmic Rbfox1 negatively regulates calcium response within the germline. We anticipate that these efforts will improve our knowledge of the human RBFOX1 function and possibly its role in human pathogenesis.