Gain-of-function screen identifies the role of miR-2 in mitochondrial homeostasis in Drosophila
Gain-of-function screen identifies the role of miR-2 in mitochondrial homeostasis in Drosophila
复制标题
功能获得筛选确定 miR-2 在果蝇线粒体稳态中的作用
DOI：
10.16380/j.kcxb.2020.11.003
复制
发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Zhang Junzheng
中科院分区：
文献类型：
--
作者：
Li Haomiao;Zhao Meiqi;Zhang Fengchao;Shen Jie;Zhang Junzheng
[Aim]真核细胞内线粒体稳态由多条分子通路共同调控，其中miRNA的重要性在最近日益凸显。最近，微小RNA（miRNAs）作为线粒体稳态的重要参与者出现了。但是对miRNA如何调控线粒体稳态仍缺乏全面准确的认识。本研究旨在探究miRNA对线粒体稳态的调控作用及机制。【方法】在果蝇中进行功能获得性筛选，在筛选过程中，miRNA在果蝇幼虫脂肪体细胞中过表达，并监测线粒体完整性。通过生物信息学方法预测参与调控线粒体稳态的miRNA下游靶标，并通过RNAi敲低实验检验靶标基因对线粒体形态的影响。【结果】共筛选到106个miRNA。其中21个miRNA的过表达导致了黑腹果蝇脂肪体的发育缺陷。当在D.在黑腹果蝇中，10个miRNA在幼虫早期致死，11个miRNA在蛹期致死。miR-2的过表达可导致果蝇幼虫脂肪体细胞线粒体异常聚集。黑腹果蝇序列分析表明，Pink 1基因可能是miR-2的靶基因。miR-2的过表达可下调Pink 1基因的表达。基因相互作用实验表明，Pink 1的过表达足以拯救miR-2引起的线粒体异常聚集。【结论】miR-2可能通过靶向Pink 1调节线粒体稳态。
【Aim】 The homeostasis of mitochondria is maintained by a wide range of molecular processes in eukaryotic cells. Very recently, microRNAs(miRNAs) are emerging as vital players of mitochondrial homeostasis. However, our understanding of how miRNAs regulate mitochondrial homeostasis is still incomplete. Our study aims to examine the roles and mechanisms of miRNAs in mitochondrial homeostasis regulation. 【Methods】 A gain-of-function screen was performed in Drosophila melanogaster during which miRNAs were over-expressed in the larval fat body cells and the mitochondrial integrity was monitored. The targets of miRNAs were predicted by bioinformatics methods and RNAi knock-down experiments were performed to examine their effects on mitochondrial morphology. 【Results】 A total of 106 miRNAs were screened in larval fat body of D. melanogaster, among which the over-expression of 21 miRNAs resulted in developmental defects of fat body. When over-expressed in the larval fat tissue of D. melanogaster, 10 miRNAs led to lethality at the early larval stage, while the other 11 miRNAs led to pupal lethality. Over-expression of miR-2 was found to result in abnormal aggregation of mitochondria in the larval fat body cells of D. melanogaster. Sequence analysis revealed that the Pink1 gene may be a target of miR-2. The expression level of Pink1 gene was shown to be down-regulated by miR-2 over-expression. Genetic interaction experiments demonstrated that over-expression of Pink1 was sufficient to rescue the abnormal aggregation of mitochondria caused by miR-2. 【Conclusion】 Our data support the view that miR-2 likely targets Pink1 to modulate mitochondrial homeostasis.