Institut de Génétique Moléculaire de Montpellier, Univ Montpellier, CNRS, Montpellier 34293, France.
Centre de Recherche en Myologie (CRM), Institut de Myologie, Sorbonne Universités, UPMC Univ Paris 06, Inserm UMRS974, GH Pitié Salpêtrière, Paris 75013, France.
RNA. 2022 Mar;28(3):303-319. doi: 10.1261/rna.078329.120. Epub 2021 Dec 10.
Spinal muscular atrophy (SMA) is a devastating neurodegenerative disease caused by reduced amounts of the ubiquitously expressed Survival of Motor Neuron (SMN) protein. In agreement with its crucial role in the biogenesis of spliceosomal snRNPs, SMN-deficiency is correlated to numerous splicing alterations in patient cells and various tissues of SMA mouse models. Among the snRNPs whose assembly is impacted by SMN-deficiency, those involved in the minor spliceosome are particularly affected. Importantly, splicing of several, but not all U12-dependent introns has been shown to be affected in different SMA models. Here, we have investigated the molecular determinants of this differential splicing in spinal cords from SMA mice. We show that the branchpoint sequence (BPS) is a key element controlling splicing efficiency of minor introns. Unexpectedly, splicing of several minor introns with suboptimal BPS is not affected in SMA mice. Using in vitro splicing experiments and oligonucleotides targeting minor or major snRNAs, we show for the first time that splicing of these introns involves both the minor and major machineries. Our results strongly suggest that splicing of a subset of minor introns is not affected in SMA mice because components of the major spliceosome compensate for the loss of minor splicing activity.
脊髓性肌萎缩症(SMA)是一种由运动神经元存活(SMN)蛋白表达量减少引起的破坏性神经退行性疾病。与 SMN 缺乏与患者细胞和 SMA 小鼠模型的各种组织中的许多剪接改变相关一致,SMN 缺乏与许多剪接体 snRNPs 的组装受到影响有关。在受 SMN 缺乏影响的 snRNPs 中,那些参与小核核糖核蛋白体的 snRNPs 受到的影响特别大。重要的是,已经表明,在不同的 SMA 模型中,几种但不是所有的 U12 依赖性内含子的剪接受到影响。在这里,我们研究了 SMA 小鼠脊髓中这种差异剪接的分子决定因素。我们表明,分支点序列(BPS)是控制小内含子剪接效率的关键因素。出乎意料的是,SMA 小鼠中几个具有次优 BPS 的小内含子的剪接不受影响。使用体外剪接实验和针对小或大 snRNA 的寡核苷酸,我们首次表明,这些内含子的剪接涉及小和大机器。我们的结果强烈表明,由于主要剪接体的成分补偿了小剪接活性的丧失,因此 SMA 小鼠中一组小内含子的剪接不受影响。