NOVA1 promotes SMN2 exon 7 splicing by binding the UCAC motif and increases SMN protein expression.
NOVA1 promotes SMN2 exon 7 splicing by binding the UCAC motif and increases SMN protein expression.
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DOI：
10.4103/1673-5374.339005
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发表时间：
2022-11
影响因子：
6.1
通讯作者：
Wu, Liu-Cheng
中科院分区：
文献类型：
--
作者：
Du, Li-Li;Sun, Jun-Jie;Chen, Zhi-Heng;Shao, Yi-Xiang;Wu, Liu-Cheng
关键词：
脊髓性肌萎缩症（SMA）是一种罕见的遗传性神经肌肉疾病，在婴儿中具有很高的致死率。据报道，运动神经元存活（SMN）1和SMN2同源基因的变异是SMA的致病因素。先前的研究表明，SMN2 外显子 7 的高包含率会增加 SMN 的表达，从而降低 SMA 的严重程度。 SMN2外显子7的包含率在神经组织中高于非神经组织。神经肿瘤腹侧抗原（NOVA）是一种在神经元中特异性且高度表达的剪接因子。它在神经系统发育和神经系统疾病的诱发中起着关键作用。然而，目前尚不清楚该剪接因子是否影响 SMA。在本研究中，我们分析了 SMA 小鼠模型（基因型 smn–/–SMN22tg/0）和同窝对照小鼠（基因型 smn+/–SMN22tg/0）不同组织中 SMN2 外显子 7 的包含情况。我们发现SMN2外显子7的包含水平在脑和脊髓组织中较高，并且NOVA1在神经系统组织中也高表达。此外，SMN2外显子7和NOVA1在中枢神经系统中同步表达。我们进一步研究了NOVA1对疾病的影响，发现SMA小鼠模型在出生后1-7天脊髓前角神经元数量减少，运动神经元中NOVA1表达水平随着脊髓性肌萎缩症的发展同时下降。我们还发现，NOVA1 的体外表达增加了 U87MG 细胞系中 SMN2 外显子 7 的包含和 SMN2 蛋白的表达，而当 NOVA1 被敲低时，观察到相反的情况。最后，点突变和RNA Pull-down表明SMN2外显子7中的UCAC基序在NOVA1结合和促进外显子7的包含中起着关键作用。此外，CA对于外显子7的包含比基序中Y残基的顺序更重要。总的来说，这些发现表明 NOVA1 与 SMN2 外显子 7 中的 UCAC 基序相互作用，从而增强 SMN2 中外显子 7 的包含，进而增加 SMN 蛋白的表达。
Spinal muscular atrophy (SMA) is a rare hereditary neuromuscular disease with a high lethality rate in infants. Variants in the homologous genes survival of motor neuron (SMN)1 and SMN2 have been reported to be SMA pathogenic factors. Previous studies showed that a high inclusion rate of SMN2 exon 7 increased SMN expression, which in turn reduced the severity of SMA. The inclusion rate of SMN2 exon 7 was higher in neural tissues than in non-neural tissues. Neuro-oncological ventral antigen (NOVA) is a splicing factor that is specifically and highly expressed in neurons. It plays a key role in nervous system development and in the induction of nervous system diseases. However, it remains unclear whether this splicing factor affects SMA. In this study, we analyzed the inclusion of SMN2 exon 7 in different tissues in a mouse model of SMA (genotype smn–/–SMN22tg/0) and littermate controls (genotype smn+/–SMN22tg/0). We found that inclusion level of SMN2 exon 7 was high in the brain and spinal cord tissue, and that NOVA1 was also highly expressed in nervous system tissues. In addition, SMN2 exon 7 and NOVA1 were expressed synchronously in the central nervous system. We further investigated the effects of NOVA1 on disease and found that the number of neurons in the anterior horn of spinal cord decreased in the mouse model of SMA during postnatal days 1–7, and that NOVA1 expression levels in motor neurons decreased simultaneously as spinal muscular atrophy developed. We also found that in vitro expression of NOVA1 increased the inclusion of SMN2 exon 7 and expression of the SMN2 protein in the U87MG cell line, whereas the opposite was observed when NOVA1 was knocked down. Finally, point mutation and RNA pull-down showed that the UCAC motif in SMN2 exon 7 plays a critical role in NOVA1 binding and promoting the inclusion of exon 7. Moreover, CA was more essential for the inclusion of exon 7 than the order of Y residues in the motif. Collectively, these findings indicate that NOVA1 interacts with the UCAC motif in exon 7 of SMN2, thereby enhancing inclusion of exon 7 in SMN2, which in turn increases expression of the SMN protein.
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影响因子：
64.5
作者：
LEFEBVRE, S;BURGLEN, L;MELKI, J
通讯作者：
MELKI, J
DOI：
10.1007/s00018-018-2849-1
发表时间：
2018-11
期刊：
Cellular and molecular life sciences : CMLS
影响因子：
--
作者：
Chaytow H;Huang YT;Gillingwater TH;Faller KME
通讯作者：
Faller KME
影响因子：
30.8
作者：
Lefebvre, S;Burlet, P;Melki, J
通讯作者：
Melki, J
影响因子：
--
作者：
Arnold, Eveline S;Fischbeck, Kenneth H
通讯作者：
Fischbeck, Kenneth H
影响因子：
9.8
作者：
Hua, Yimin;Vickers, Timothy A;Baker, Brenda F;Bennett, C Frank;Krainer, Adrian R
通讯作者：
Krainer, Adrian R