SRp54 (SFRS11), a regulator for tau exon 10 alternative splicing identified by an expression cloning strategy
SRp54 (SFRS11), a regulator for tau exon 10 alternative splicing identified by an expression cloning strategy
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DOI：
10.1128/mcb.00739-06
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发表时间：
2006-09-01
影响因子：
5.3
通讯作者：
Havlioglu, Necat
中科院分区：
文献类型：
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作者：
Wu, Jane Y.;Kar, Amar;Havlioglu, Necat
Tau基因编码一种微管相关蛋白，对神经元的生存和功能至关重要。人类tau基因的剪接缺陷会导致额颞部痴呆，并伴有与17号染色体相关的帕金森症(FTDP-17)，这是一种常染色体显性遗传性神经退行性疾病。与FTDP-17相关的基因突变经常影响tau外显子10的选择性剪接。为了研究tau外显子10选择性剪接的调控机制，我们开发了一个tau外显子10跳过的绿色荧光蛋白报告和一种识别剪接调控因子的表达克隆策略。使用这一策略发现了SRp54(也称为SFRS11)作为tau外显子10剪接抑制因子的作用。SRp54的过表达抑制了tau外显子10的包涵体。RNA干扰介导的SRp54基因敲除增加了外显子10的包涵体。SRp54与外显子10中的一个富含嘌呤的元件相互作用，并拮抗Tra2β，Tra2β是一种含有SR结构域的蛋白，可以增强外显子10的包含性。该外显子元件的缺失消除了SRp54抑制外显子10包涵体的活性。我们的数据支持SRp54在调节tau外显子10剪接中的作用。这些实验还建立了一种普遍有用的方法，用于通过表达克隆来识别选择性剪接的反式作用调节因子。
The tau gene encodes a microtubule-associated protein that is critical for neuronal survival and function. Splicing defects in the human tau gene lead to frontotemporal dementia with Parkinsonism linked to chromosome 17 (FTDP-17), an autosomal dominant neurodegenerative disorder. Genetic mutations associated with FTDP-17 often affect tau exon 10 alternative splicing. To investigate mechanisms regulating tau exon 10 alternative splicing, we have developed a green fluorescent protein reporter for tau exon 10 skipping and an expression cloning strategy to identify splicing regulators. A role for SRp54 (also named SFRS11) as a tau exon 10 splicing repressor has been uncovered using this strategy. The overexpression of SRp54 suppresses tau exon 10 inclusion. RNA interference-mediated knock-down of SRp54 increases exon 10 inclusion. SRp54 interacts with a purine-rich element in exon 10 and antagonizes Tra2 beta, an SR-domain-containing protein that enhances exon 10 inclusion. Deletion of this exonic element eliminates the activity of SRp54 in suppressing exon 10 inclusion. Our data support a role of SRp54 in regulating tau exon 10 splicing. These experiments also establish a generally useful approach for identifying trans-acting regulators of alternative splicing by expression cloning.