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

RNA 结合蛋白 Rbfox 的新功能的分子表征

• 批准号: 8598295
• 负责人: Arnaldo Carreira-Rosario
• 金额: $ 2.97万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8598295
• 关键词: 3' Untranslated Regions; Adult; Affect; Alternative Splicing; Autistic Disorder; Binding; Biological Models; Calcium; Calcium/calmodulin-dependent protein kinase; Calmodulin; Cell Nucleus; Cell physiology; Cells; Cytoplasm; Data; Development; Diagnostic; Disease; Drosophila genus; Elements; Epilepsy; Family; Foxes; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Translation; Genetic screening method; Goals; Human; Knowledge; Link; Malignant Neoplasms; Mammalian Cell; Measures; Mediating; Mental Retardation; Messenger RNA; Metabolism; Molecular; Molecular Genetics; Motivation; Mutation; Non-Small-Cell Lung Carcinoma; Nuclear; Oogenesis; Pathogenesis; Play; Post-Transcriptional Regulation; Process; Protein Isoforms; Proteins; RNA; RNA Binding; RNA Interference; RNA Recognition Motif; RNA Splicing; RNA-Binding Proteins; Regulation; Reporter; Resolution; Reverse Transcriptase Polymerase Chain Reaction; Role; System; Testing; Tissue-Specific Splicing; Tissues; Transcript; Transgenic Organisms; Translational Repression; Translations; Untranslated Regions; base; calmodulin-dependent protein kinase II; cell type; design; fly; gene function; improved; in vivo; insight; mRNA Instability; malignant stomach neoplasm; member; mutant; nervous system disorder; novel; public health relevance; response; sensor; therapeutic development; tool

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.

描述（由申请人提供）：转录后基因调控已成为一种重要的控制机制，在发育和成年期间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，调节一系列过程，包括剪接和翻译。一个这样的家族是RNA结合fox（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，RBFOX 1与从神经系统疾病到癌症的许多人类疾病有关。RBFOX1基因编码几种亚型：调节组织特异性选择性剪接的核亚型和胞质亚型，其功能尚不清楚。在果蝇生殖系中，Rbfox1在细胞质中观察到，设计用于特异性降低细胞质Rbfox1的RNAi导致生殖系分化受损。我们已经证明Rbfox1抑制在其3'UTR内含有（U）GCAUG元件的转录物的翻译。令人兴奋的是，我们的研究结果提出了一个新的功能，广泛研究的剪接调节器，并提供了这个建议的基础。的 该提案的主要目标是提供对RBFOX 1的分子和发育功能的更深入理解。我们的总体策略是使用果蝇特有的复杂的遗传和分子工具来研究Rbfox1在体内的功能。我们将利用我们已经分离的Rbfox1突变株系，从遗传学上研究Rbfox1控制翻译的分子机制。最后，我们将探讨细胞质Rbfox1负调控钙反应的可能性。我们预计，这些努力将提高我们对人类RBFOX 1功能的认识，并可能在人类发病机制中发挥作用。