ROLE OF ELAV IN NEURONAL RNA PROCESSING

描述(申请人提供)：这项研究项目的长期目标是了解神经元中差异表达的反式作用因子如何控制前mRNA的处理，以精确调控神经元基因的表达。利用果蝇强大的遗传学、分子生物学和转基因技术，以及新兴的DNA微阵列技术，将研究ELAV在mRNA加工中的作用及其对神经元基因表达的整体影响。ELAV是ELAV/HU RNA结合蛋白家族的创始成员，该家族在脊椎动物和无脊椎动物中都是保守的。ELAV/Hu家族的成员在mRNA加工中扮演不同的角色，包括剪接、稳定性和可译性。该项目的目的是：(1)阐明ELAV与RNA转录本相互作用的机制；(2)利用免疫沉淀ELAV-核蛋白复合体和DNA微阵列鉴定ELAV的直接靶点；(3)利用微阵列技术评估ELAV对基因表达的整体影响。总而言之，这些方法将开始识别受集体调控的基因网络，并提供如何在神经元基因表达中利用转录后调控的全面视角。鉴于果蝇和人类基因组之间的进化保守，在调控方面的见解 战略将直接转移到人类研究中。人类ELAV/Hu家族成员与副肿瘤性小脑功能障碍的发病机制有关。RNA加工缺陷在大量人类疾病和遗传性疾病中被证明存在，包括癌症、肌营养不良症和脆性X综合征。

DESCRIPTION (provided by applicant): The long-term goal of this research project is to understand how differentially-expressed trans-acting factors in neurons can control pre-mRNA processing to precisely regulate neuronal gene expression. The powerful genetics, molecular biology and transgenic techniques of Drosophila, along with the emergent DNA microarray technology, will be utilized to study the role of ELAV in mRNA processing and its overall impact on gene expression in neurons. ELAV is the founding member of the ELAV/Hu family of RNA-binding proteins, which is conserved in both vertebrates and invertebrates. Members of the ELAV/Hu family serve diverse roles in mRNA processing, including splicing, stability and translatability. The aims of the project are: (1) elucidation of mechanisms of ELAV's interactions with RNA transcripts, (2) identification of direct targets of ELAV using immunoprecipitated ELAV-ribnucleoprotein complexes and DNA microarrays, (3) assessment of the overall impact of ELAV on gene expression using microarray technology. Together, these approaches will begin to identify networks of genes that are regulated collectively and provide a comprehensive view of how post-transcriptional regulation is utilized in neuronal gene expression. Given the evolutionary conservation between Drosophila and human genomes, insights in regulatory strategies will be directly transferable to human studies. Human members of the ELAV/Hu family have been implicated in pathogenesis of paraneoplastic cerebellar dysfunction. RNA processing defects have been documented in a large number of human diseases and inherited disorders including cancer, muscular dystrophy and fragile X syndrome.