Wijegunawardana Denethi, Vishal Sonali S, Venkatesh Neha, Gopal Pallavi P
bioRxiv. 2023 Feb 15:2023.01.30.526372. doi: 10.1101/2023.01.30.526372.
Altered RNA metabolism is a common pathogenic mechanism linked to familial and sporadic Amyotrophic lateral sclerosis (ALS). ALS is characterized by mislocalization and aggregation of TDP-43, an RNA-binding protein (RBP) with multiple roles in post-transcriptional RNA processing. Recent studies have identified genetic interactions between TDP-43 and Ataxin-2, a polyglutamine (polyQ) RBP in which intermediate length polyQ expansions confer increased ALS risk. Here, we used live-cell confocal imaging, photobleaching and translation reporter assays to study the localization, transport dynamics and mRNA regulatory functions of TDP-43/Ataxin-2 in rodent primary cortical neurons. We show that Ataxin-2 polyQ expansions aberrantly sequester TDP-43 within ribonucleoprotein (RNP) condensates, and disrupt both its motility along the axon and liquid-like properties. Our data suggest that Ataxin-2 governs motility and translation of neuronal RNP condensates and that Ataxin-2 polyQ expansions fundamentally perturb spatial localization of mRNA and suppress local translation. Overall, these results indicate Ataxin-2 polyQ expansions have detrimental effects on stability, localization, and translation of transcripts critical for axonal and cytoskeletal integrity, particularly important for motor neurons.
RNA代谢改变是一种与家族性和散发性肌萎缩侧索硬化症(ALS)相关的常见致病机制。ALS的特征是TDP-43的错误定位和聚集,TDP-43是一种在转录后RNA加工中具有多种作用的RNA结合蛋白(RBP)。最近的研究确定了TDP-43与Ataxin-2之间的遗传相互作用,Ataxin-2是一种多聚谷氨酰胺(polyQ)RBP,其中等长度的polyQ扩展会增加患ALS的风险。在这里,我们使用活细胞共聚焦成像、光漂白和翻译报告分析来研究TDP-43/Ataxin-2在啮齿动物原代皮质神经元中的定位、转移动力学和mRNA调节功能。我们发现Ataxin-2的polyQ扩展异常地将TDP-43隔离在核糖核蛋白(RNP)凝聚物中,并破坏其沿轴突的运动性和类液体特性。我们的数据表明,Ataxin-2控制神经元RNP凝聚物的运动性和翻译,并且Ataxin-2的polyQ扩展从根本上扰乱mRNA的空间定位并抑制局部翻译。总体而言,这些结果表明Ataxin-2的polyQ扩展对轴突和细胞骨架完整性至关重要的转录本的稳定性、定位和翻译具有有害影响,这对运动神经元尤为重要。