Novel roles of Drosophila FUS and Aub responsible for piRNA biogenesis in neuronal disorders.
Novel roles of Drosophila FUS and Aub responsible for piRNA biogenesis in neuronal disorders.
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果蝇 FUS 和 Aub 在神经元疾病中负责 piRNA 生物发生的新作用。
DOI：
10.1016/j.brainres.2018.12.028
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发表时间：
2018
期刊：
影响因子：
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通讯作者：
Yamaguchi M.
中科院分区：
文献类型：
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作者：
Wakisaka KT;Tanaka R;Hirashima T;Muraoka Y;Azuma Y;Yoshida H;Tokuda T;Asada S;Suda K;Ichiyanagi K;Ohno S;Itoh M;Yamaguchi M.
PiRNAs是一种小的非编码RNA，被认为仅限于生殖细胞。尽管最近在包括神经元在内的体细胞中检测到了它们，但目前还不清楚piRNA的生物发生如何参与神经元疾病。在这里，我们使用Cabeza(Caz)击倒果蝇，研究了Aubergine(Aub)，一种负责piRNA生物发生的Piwi家族蛋白(PIWI)在神经元疾病中的可能作用。Cazis是FUS的果蝇同源物，FUS是导致肌萎缩侧索硬化症(ALS)的基因之一。AUB的过表达增加了神经元特异性Caz基因敲除所致的神经肌肉接头运动神经元的运动神经元运动缺陷和解剖缺陷。为了阐明潜在的机制，我们检测了在这些条件下的前-piRNA和成熟大小的piRNA水平。QRT-PCR和RNA-seq分析表明，Caz基因敲除增加了中枢神经系统(CNS)中前-piRNA的水平，但降低了成熟大小的piRNA水平，这表明它在前-piRNA的产生中发挥了作用。AUB过表达并不增加成熟大小的piRNA水平。这些结果表明，积累的前piRNAs是异常流产的prepiRNAs，不能被切片器进一步处理，包括Aub。我们还通过RNA免疫沉淀证明了CAZ和CNS中的前piRNAs之间的关系。AUB过表达导致CAZ胞浆定位异常。在这些结果的基础上，我们提出了一个模型，在该模型中，Caz基因击倒导致异常的Pre-piRNAs与Caz相关，然后在细胞质中移位和积聚，这一过程可能由Aub介导。本文中使用Caz基因敲除的Fly所展示的CAZ和AUB的新角色将有助于更深入地理解ALS的发病机制。
piRNAs, small non-coding RNAs, were considered to be restricted to germline cells. Although they have recently been detected in somatic cells including neurons, it remains unclear how piRNA biogenesis is involved in neuronal diseases. We herein examined the possible roles of Aubergine (Aub), a Piwi-family protein (PIWI) responsible for piRNA biogenesis, in the neuronal disorders, using theCabeza(Caz) knockdownDrosophila. Cazis aDrosophilahomologue ofFUS, which is one of the genes causing amyotrophic lateral sclerosis (ALS). Aub overexpression enhanced the mobility defects accompanied by anatomical defects in motoneurons at neuromuscular junctions induced by the neuron-specific knockdown ofCaz. In order to elucidate the underlying mechanisms, we examined pre-piRNA and mature-size piRNA levels under these conditions. qRT-PCR and RNA-seq analyses revealed that theCazknockdown increased pre-piRNA levels, but reduced mature-size piRNA levels in the central nervous system (CNS), suggesting a role in the pre-piRNAs production. Aub overexpression did not increase mature-size piRNA levels. These results suggest that the accumulated pre-piRNAs are abnormal abortive pre-piRNAs that cannot be further processed by slicers, including Aub. We also demonstrated a relationship between Caz and pre-piRNAs in the CNS by RNA immunoprecipitation. Aub overexpression induced the abnormal cytoplasmic localization of Caz. Based on these results, we propose a model in whichCazknockdown-induced abnormal pre-piRNAs associate with Caz, then translocate and accumulate in the cytoplasm, a process that may be mediated by Aub. The novel roles for Caz and Aub demonstrated herein using theCaz-knockdown fly will contribute to a deeper understanding of the pathogenesis of ALS.