Comparative interactomics analysis of different ALS-associated proteins identifies converging molecular pathways.
Comparative interactomics analysis of different ALS-associated proteins identifies converging molecular pathways.
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DOI：
10.1007/s00401-016-1575-8
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发表时间：
2016-08
影响因子：
12.7
通讯作者：
Pasterkamp RJ
中科院分区：
文献类型：
--
作者：
Blokhuis AM;Koppers M;Groen EJN;van den Heuvel DMA;Dini Modigliani S;Anink JJ;Fumoto K;van Diggelen F;Snelting A;Sodaar P;Verheijen BM;Demmers JAA;Veldink JH;Aronica E;Bozzoni I;den Hertog J;van den Berg LH;Pasterkamp RJ
肌萎缩侧索硬化症（ALS）是一种毁灭性的神经系统疾病，目前尚无有效的治疗方法。越来越多的 ALS 遗传原因正在被确定，但这些遗传缺陷如何导致运动神经元变性以及它们在多大程度上影响常见的细胞通路仍不完全清楚。为了解决这些问题，我们进行了相互作用组学分析，以确定神经元细胞中 ATXN2、C9orf72、FUS、OPTN、TDP-43 和 UBQLN2 的野生型 (WT) 和 ALS 相关突变版本的结合伴侣。该分析鉴定了这些蛋白质的几种已知的但也有许多新的结合配偶体。 WT 和突变 ALS 蛋白的相互作用组非常相似，但 OPTN 和 UBQLN2 除外，其中突变导致蛋白质相互作用的丧失或增强。几个已确定的相互作用组表现出高度重叠：ATXN2、FUS 和 TDP-43 的共享结合伙伴在 RNA 代谢中发挥作用； OPTN 和 UBQLN2 相互作用蛋白与线粒体中的蛋白质降解和蛋白质转运以及 C9orf72 相互作用蛋白功能相关。为了证实这种重叠对于 ALS 发病机制很重要，我们在 FUS ALS 的体外和体内模型系统中更详细地研究了脆性 X 智力迟钝蛋白 (FMRP)，它是 ATXN2、FUS 和 TDP-43 的常见相互作用因子之一。 FMRP 定位于脊髓运动神经元中含有突变 FUS 的聚集体，并以直接且 RNA 敏感的方式结合内源性 FUS。此外，斑马鱼胚胎中突变 FUS 引起的突触 FMRP mRNA 靶标表达、神经肌肉接头完整性和运动行为缺陷可以通过外源 FMRP 表达来挽救。总之，这些结果表明，相互作用组学分析可以为 ALS 疾病机制提供重要的见解，并将 FMRP 与 FUS 突变引起的运动神经元功能障碍联系起来。本文的在线版本 (doi:10.1007/s00401-016-1575-8) 包含补充材料，可供授权用户使用。
Amyotrophic lateral sclerosis (ALS) is a devastating neurological disease with no effective treatment available. An increasing number of genetic causes of ALS are being identified, but how these genetic defects lead to motor neuron degeneration and to which extent they affect common cellular pathways remains incompletely understood. To address these questions, we performed an interactomic analysis to identify binding partners of wild-type (WT) and ALS-associated mutant versions of ATXN2, C9orf72, FUS, OPTN, TDP-43 and UBQLN2 in neuronal cells. This analysis identified several known but also many novel binding partners of these proteins. Interactomes of WT and mutant ALS proteins were very similar except for OPTN and UBQLN2, in which mutations caused loss or gain of protein interactions. Several of the identified interactomes showed a high degree of overlap: shared binding partners of ATXN2, FUS and TDP-43 had roles in RNA metabolism; OPTN- and UBQLN2-interacting proteins were related to protein degradation and protein transport, and C9orf72 interactors function in mitochondria. To confirm that this overlap is important for ALS pathogenesis, we studied fragile X mental retardation protein (FMRP), one of the common interactors of ATXN2, FUS and TDP-43, in more detail in in vitro and in vivo model systems for FUS ALS. FMRP localized to mutant FUS-containing aggregates in spinal motor neurons and bound endogenous FUS in a direct and RNA-sensitive manner. Furthermore, defects in synaptic FMRP mRNA target expression, neuromuscular junction integrity, and motor behavior caused by mutant FUS in zebrafish embryos, could be rescued by exogenous FMRP expression. Together, these results show that interactomics analysis can provide crucial insight into ALS disease mechanisms and they link FMRP to motor neuron dysfunction caused by FUS mutations. The online version of this article (doi:10.1007/s00401-016-1575-8) contains supplementary material, which is available to authorized users.