Munich Cluster for Systems Neurology, Medical Faculty, Ludwig-Maximilians-University München, Munich, Germany.
Department of Biology, University of Fribourg, Fribourg, Switzerland.
Life Sci Alliance. 2022 Jul 1;5(11). doi: 10.26508/lsa.202101327. Print 2022 Nov.
Ubiquilin-2 (UBQLN2) is a ubiquitin-binding protein that shuttles ubiquitinated proteins to proteasomal and autophagic degradation. UBQLN2 mutations are genetically linked to the neurodegenerative disorders amyotrophic lateral sclerosis and frontotemporal dementia (ALS/FTD). However, it remains elusive how UBQLN2 mutations cause ALS/FTD. Here, we systematically examined proteomic and transcriptomic changes in patient-derived lymphoblasts and CRISPR/Cas9-engineered HeLa cells carrying ALS/FTD UBQLN2 mutations. This analysis revealed a strong up-regulation of the microtubule-associated protein 1B (MAP1B) which was also observed in UBQLN2 knockout cells and primary rodent neurons depleted of UBQLN2, suggesting that a UBQLN2 loss-of-function mechanism is responsible for the elevated MAP1B levels. Consistent with MAP1B's role in microtubule binding, we detected an increase in total and acetylated tubulin. Furthermore, we uncovered that UBQLN2 mutations result in decreased phosphorylation of MAP1B and of the ALS/FTD-linked fused in sarcoma (FUS) protein at S439 which is critical for regulating FUS-RNA binding and MAP1B protein abundance. Together, our findings point to a deregulated UBQLN2-FUS-MAP1B axis that may link protein homeostasis, RNA metabolism, and cytoskeleton dynamics, three molecular pathomechanisms of ALS/FTD.
泛素结合蛋白 2(UBQLN2)是一种泛素结合蛋白,可将泛素化蛋白转运至蛋白酶体和自噬降解。UBQLN2 突变与神经退行性疾病肌萎缩侧索硬化症和额颞叶痴呆(ALS/FTD)有遗传关联。然而,UBQLN2 突变如何导致 ALS/FTD 仍不清楚。在这里,我们系统地研究了携带 ALS/FTD UBQLN2 突变的患者来源的淋巴母细胞和 CRISPR/Cas9 工程化的 HeLa 细胞中的蛋白质组和转录组变化。这项分析显示微管相关蛋白 1B(MAP1B)的强烈上调,这在 UBQLN2 敲除细胞和耗尽 UBQLN2 的原代啮齿动物神经元中也观察到,这表明 UBQLN2 功能丧失机制负责升高的 MAP1B 水平。与 MAP1B 在微管结合中的作用一致,我们检测到总和乙酰化微管蛋白的增加。此外,我们发现 UBQLN2 突变导致 MAP1B 和融合肉瘤(FUS)蛋白的 S439 处的磷酸化减少,这对于调节 FUS-RNA 结合和 MAP1B 蛋白丰度至关重要。总之,我们的研究结果表明,UBQLN2-FUS-MAP1B 轴失调可能与蛋白质稳态、RNA 代谢和细胞骨架动力学有关,这是 ALS/FTD 的三个分子发病机制。