FUS ALS-causative mutations impair FUS autoregulation and splicing factor networks through intron retention
FUS ALS-causative mutations impair FUS autoregulation and splicing factor networks through intron retention
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DOI：
10.1093/nar/gkaa410
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发表时间：
2020-07-09
影响因子：
14.9
通讯作者：
Fratta, Pietro
中科院分区：
文献类型：
--
作者：
Humphrey, Jack;Birsa, Nicol;Fratta, Pietro
RNA 结合蛋白 FUS 的突变会导致肌萎缩侧索硬化症 (ALS)，这是一种毁灭性的神经退行性疾病。 FUS 在 RNA 代谢的许多方面发挥着作用，包括 mRNA 剪接。然而，ALS 致病突变对剪接的影响尚未得到充分表征，因为大多数疾病模型都是基于过表达突变型 FUS，这会由于 FUS 自动调节而改变 RNA 加工。我们和其他人最近创建了克服过度表达问题的敲入模型，并在 FUS 敲除的同时对 FUS 突变体进行了深度 RNA 测序，使我们能够将突变引起的变化与真正的功能丧失进行比较。我们发现 FUS-ALS 突变会导致表达和剪接功能广泛丧失。具体来说，我们发现突变的 FUS 直接改变 RNA 结合蛋白的内含子保留水平。此外，我们还发现 FUS 本身存在与其自动调节相关的内含子保留事件。 FUS 水平的改变与疾病有关，我们在此表明这种新的自动调节机制会因 FUS 突变而改变。至关重要的是，我们还在其他遗传形式的 ALS 中观察到了这种现象，包括由 TDP-43、VCP 和 SOD1 突变引起的那些，支持了多个 ALS 基因在调控网络中相互作用的概念。
Mutations in the RNA-binding protein FUS cause amyotrophic lateral sclerosis (ALS), a devastating neurodegenerative disease. FUS plays a role in numerous aspects of RNA metabolism, including mRNA splicing. However, the impact of ALS-causative mutations on splicing has not been fully characterized, as most disease models have been based on overexpressing mutant FUS, which will alter RNA processing due to FUS autoregulation. We and others have recently created knockin models that overcome the overexpression problem, and have generated high depth RNA-sequencing on FUS mutants in parallel to FUS knockout, allowing us to compare mutation-induced changes to genuine loss of function. We find that FUS-ALS mutations induce a widespread loss of function on expression and splicing. Specifically, we find that mutant FUS directly alters intron retention levels in RNA-binding proteins. Moreover, we identify an intron retention event in FUS itself that is associated with its autoregulation. Altered FUS levels have been linked to disease, and we show here that this novel autoregulation mechanism is altered by FUS mutations. Crucially, we also observe this phenomenon in other genetic forms of ALS, including those caused by TDP-43, VCP and SOD1 mutations, supporting the concept that multiple ALS genes interact in a regulatory network.