Mechanisms regulating tau alternative pre-mRNA splicing

描述（由申请人提供）：调节tau替代前mRNA剪接的机制累积的证据表明，前mRNA剪接缺陷和异常剪接导致一系列神经退行性疾病。选择性剪接对tau基因功能至关重要，人类tau基因中的剪接突变导致额颞叶痴呆（FTLD-tau）。虽然tau剪接突变最初于1998年报道，但突变影响tau选择性剪接的机制仍然知之甚少，FTLD-tau中的风险因素以及遗传修饰剂仍有待确定。本研究旨在探讨tau基因选择性剪接的调控机制。我们计划使用分子，生物化学和细胞生物学方法来研究tau蛋白的选择性剪接，并比较野生型与突变型tau蛋白转录本的差异剪接体识别。 我们已经建立了体外生化检测和细胞培养系统来研究tau外显子10的选择性剪接。使用生化分析和表达克隆策略，我们已经确定了几个蛋白参与tau外显子10选择性剪接在我们的初步研究。还建立了原代神经元培养系统来研究已确定的因素在神经元细胞死亡中的作用。对tau蛋白选择性剪接的顺式和反式作用元件的分子解剖不仅有助于理解控制tau蛋白选择性剪接的基本机制，而且还可以揭示神经退行性变发病机制中的新参与者。我们计划采用生物信息学、生物化学、分子和遗传学相结合的方法来研究FTLD-tau蛋白的发病机制，这是一种最常见的神经退行性疾病。

DESCRIPTION (provided by applicant): Mechanisms regulating tau alternative pre-mRNA splicing Accumulating evidence indicates that pre-mRNA splicing defects and aberrant splicing lead to a range of neurodegenerative disorders. Alternative splicing is critical for tau gene function, and splicing mutations in the human tau gene lead to fronto-temporal lobe dementia (FTLD-tau). Although tau splicing mutations were initially reported in 1998, the mechanisms by which mutations affect tau alternative splicing are still poorly understood, and risk factors as well as genetic modifiers in FTLD-tau remain to be identified. This proposal aims to investigate the mechanisms regulating tau gene alternative splicing. We plan to use molecular, biochemical and cell biological methods to investigate tau alternative splicing and to compare the differential spliceosomal recognition of wild type versus mutant tau transcripts. We have established both in vitro biochemical assays and cell culture systems to investigate tau exon 10 alternative splicing. Using biochemical assays and an expression cloning strategy, we have identified several proteins involved in tau exon 10 alternative splicing in our preliminary studies. A primary neuronal culture system has also been set up to investigate the role of identified factors in neuronal cell death. Molecular dissection of cis- and trans-acting elements important for tau alternative splicing will not only help in understanding the basic mechanisms controlling tau alternative splicing but also reveal new players in pathogenesis of neurodegeneration. We plan to use combined bioinformatics, biochemical, molecular and genetic approaches to study the pathogenetic mechanisms underlying FTLD-tau, one most common neurodegenerative disorders.