ALS-linked FUS mutations dysregulate G-quadruplex-dependent liquid-liquid phase separation and liquid-to-solid transition.
ALS-linked FUS mutations dysregulate G-quadruplex-dependent liquid-liquid phase separation and liquid-to-solid transition.
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DOI：
10.1016/j.jbc.2021.101284
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发表时间：
2021-11
期刊：
影响因子：
--
通讯作者：
Ishihama A
中科院分区：
文献类型：
--
作者：
Ishiguro A;Lu J;Ozawa D;Nagai Y;Ishihama A
肌萎缩侧索硬化症（amyotrophiclateralsclerosis，ALS）是一种以运动神经元内蛋白质聚集为特征的神经退行性疾病。最近在ALS患者中发现的基因突变促进了对ALS潜在的复杂分子机制的研究。FUS（融合在肉瘤中）是一种代表性的ALS连接的RNA结合蛋白（RBP），其特异性识别G-四链体（G4）-DNA/RNA。然而，ALS连锁的FUS突变对G4-RNA结合活性和相行为的影响从未被研究过。使用纯化的全长FUS，我们分析了由多个功能模块组成的多结构域结构与G4结合的分子机制。在这里，我们成功地观察到FUS冷凝物形成的液-液相分离（LLPS）和随后的液-固转变（LST），导致FUS聚集体的形成。该过程通过FUS与G4-RNA的相互作用而显著促进。为了进一步研究，我们选择了总共8个代表性ALS连锁FUS突变体内的多结构域结构和纯化这些蛋白质。G4-RNA依赖性LLPS和LST途径的调节在所有ALS连锁的FUS突变体中丢失，这些突变体在测试的G4-RNA识别中有缺陷，这支持了G4-RNA在该过程中的重要作用。值得注意的是，导致幼年型ALS的P525L突变对G4-RNA结合和FUS聚集的影响最大。本文所述的发现可以为ALS发病机制的复杂途径中蛋白质聚集和RBP功能障碍之间迄今未定义的联系提供线索。
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by the accumulation of protein aggregates in motor neurons. Recent discoveries of genetic mutations in ALS patients promoted research into the complex molecular mechanisms underlying ALS. FUS (fused in sarcoma) is a representative ALS-linked RNA-binding protein (RBP) that specifically recognizes G-quadruplex (G4)-DNA/RNAs. However, the effects of ALS-linked FUS mutations on the G4-RNA-binding activity and the phase behavior have never been investigated. Using the purified full-length FUS, we analyzed the molecular mechanisms of multidomain structures consisting of multiple functional modules that bind to G4. Here we succeeded to observe the liquid–liquid phase separation (LLPS) of FUS condensate formation and subsequent liquid-to-solid transition (LST) leading to the formation of FUS aggregates. This process was markedly promoted through FUS interaction with G4-RNA. To further investigate, we selected a total of eight representative ALS-linked FUS mutants within multidomain structures and purified these proteins. The regulation of G4-RNA-dependent LLPS and LST pathways was lost for all ALS-linked FUS mutants defective in G4-RNA recognition tested, supporting the essential role of G4-RNA in this process. Noteworthy, the P525L mutation that causes juvenile ALS exhibited the largest effect on both G4-RNA binding and FUS aggregation. The findings described herein could provide a clue to the hitherto undefined connection between protein aggregation and dysfunction of RBPs in the complex pathway of ALS pathogenesis.
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影响因子：
2.1
作者：
Ishiguro, Akira;Kimura, Nobuyuki;Ishihama, Akira
通讯作者：
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15.1
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5.6
作者：
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通讯作者：
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