Alternative splicing regulation of APP exon 7 by RBFox proteins
Alternative splicing regulation of APP exon 7 by RBFox proteins
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DOI：
10.1016/j.neuint.2014.08.001
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发表时间：
2014-12-01
影响因子：
4.2
通讯作者：
Tsukahara, Toshifumi
中科院分区：
文献类型：
--
作者：
Alam, Shafiul;Suzuki, Hitoshi;Tsukahara, Toshifumi
RBFox蛋白是众所周知的选择性剪接调节因子。我们以前已经证明，在全反式维甲酸诱导P19细胞分化和细胞聚集的过程中，RBFOX1的时间表达显著增加。为了找到其关键的剪接调控，我们检查了RBFOX1对33个先前报道和验证的P19细胞神经元剪接事件的影响。我们观察到，在HEK293和HeLa细胞中，瞬时表达RBFOX1改变了三个基因的选择性剪接，特别是淀粉样前体蛋白(APP)、盘状大同系物3(Dlg3)和G蛋白(α激活活性多肽O(GNAO1))。此外，不能与靶RNA序列((U)GCAUG)结合的RBFOX1突变体(RBFOX1 FA)不能诱导这些剪接事件。APP产生淀粉样β蛋白多肽，参与阿尔茨海默病的病理，因此我们研究了RBFOX1和其他剪接调节因子对APP的替代剪接调控。我们的结果表明，RBFox蛋白促进APP外显子7的跳过，但不包括外显子8。我们制作了APP6789微型基因，观察到位于外显子7上游和外显子7的两个(U)GCAUG序列具有诱导RBFox蛋白跳过外显子7的功能。总体而言，RBFox蛋白可能会将APP从含有APP770和APP751的外显子7转移到缺少APP695的外显子7，APP695在神经组织中占主导地位。(C)2014爱思唯尔有限公司。保留所有权利。
RBFox proteins are well-known alternative splicing regulators. We have shown previously that during neuronal differentiation of P19 cells induced by all-trans retinoic acid and cell aggregation, RBFox1 shows markedly increased temporal expression. To find its key splicing regulation, we examined the effect of RBFox1 on 33 previously reported and validated neuronal splicing events of P19 cells. We observed that alternative splicing of three genes, specifically, amyloid precursor protein (APP), disks large homolog 3 (DLG3), and G protein, alpha activating activity polypeptide O (GNAO1), was altered by transient RBFox1 expression in HEK293 and HeLa cells. Moreover, an RBFox1 mutant (RBFox1 FA) that was unable to bind the target RNA sequence ((U)GCAUG) did not induce these splicing events. APP generates amyloid beta peptides that are involved in the pathology of Alzheimer's disease, and therefore we examined APP alternative splicing regulation by RBFox1 and other splicing regulators. Our results indicated that RBFox proteins promote the skipping of APP exon 7, but not the inclusion of exon 8. We made APP6789 minigenes and observed that two (U)GCAUG sequences, located upstream of exon 7 and in exon 7, functioned to induce skipping of exon 7 by RBFox proteins. Overall, RBFox proteins may shift APP from exon 7 containing isoforms, APP770 and APP751, toward the exon 7 lacking isoform, APP695, which is predominant in neural tissues. (C) 2014 Elsevier Ltd. All rights reserved.