The 3'-5' exoribonuclease Dis3 regulates the expression of specific microRNAs in Drosophila wing imaginal discs.
The 3'-5' exoribonuclease Dis3 regulates the expression of specific microRNAs in Drosophila wing imaginal discs.
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DOI：
10.1080/15476286.2015.1040978
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发表时间：
2015
期刊：
影响因子：
4.1
通讯作者：
Newbury SF
中科院分区：
文献类型：
--
作者：
Towler BP;Jones CI;Viegas SC;Apura P;Waldron JA;Smalley SK;Arraiano CM;Newbury SF
Dis 3是一种高度保守的核糖核酸外切酶，其在3 '-5'方向降解RNA。Dis 3的突变与许多人类癌症有关，包括多发性骨髓瘤和急性髓性白血病。在这项工作中，我们评估了一个Dis 3敲低果蝇成虫盘发育和成熟microRNA的表达的影响。我们发现，Dis 3基因敲低严重破坏了翅膀成虫盘的发育，苍蝇有一个“无翼”表型。使用RNA-seq定量Dis 3敲低对microRNA表达的影响表明，Dis 3通常调节一小部分microRNA，只有11个（10.1%）水平增加≥2倍，6个（5.5%）水平降低≥2倍。在这些microRNA中，与对照相比，miR-252- 5 p在Dis 3耗尽的细胞中增加2.1倍，而miR-252前体的水平不变，这表明Dis 3可以在细胞质中起作用以特异性降解这种成熟的miRNA。此外，我们的实验表明，Dis 3通常与细胞质中的外泌体亚基Rrp 40相互作用，以靶向miR-252- 5 p，从而在正常翅膀发育期间降解。另一种microRNA，miR-982- 5 p，在Dis 3敲低细胞中以较低水平表达，而miR-982前体保持不变，表明Dis 3参与其加工。因此，我们的研究揭示了这种核糖核酸酶对microRNA调控的意想不到的特异性，这可能在其他真核生物中是保守的，并且可能与理解其在人类疾病中的作用有关。
Dis3 is a highly conserved exoribonuclease which degrades RNAs in the 3'-5' direction. Mutations in Dis3 are associated with a number of human cancers including multiple myeloma and acute myeloid leukemia. In this work, we have assessed the effect of a Dis3 knockdown on Drosophila imaginal disc development and on expression of mature microRNAs. We find that Dis3 knockdown severely disrupts the development of wing imaginal discs in that the flies have a “no wing” phenotype. Use of RNA-seq to quantify the effect of Dis3 knockdown on microRNA expression shows that Dis3 normally regulates a small subset of microRNAs, with only 11 (10.1%) increasing in level ≥2-fold and 6 (5.5%) decreasing in level ≥2-fold. Of these microRNAs, miR-252–5p is increased 2.1-fold in Dis3-depleted cells compared to controls while the level of the miR-252 precursor is unchanged, suggesting that Dis3 can act in the cytoplasm to specifically degrade this mature miRNA. Furthermore, our experiments suggest that Dis3 normally interacts with the exosomal subunit Rrp40 in the cytoplasm to target miR-252–5p for degradation during normal wing development. Another microRNA, miR-982–5p, is expressed at lower levels in Dis3 knockdown cells, while the miR-982 precursor remains unchanged, indicating that Dis3 is involved in its processing. Our study therefore reveals an unexpected specificity for this ribonuclease toward microRNA regulation, which is likely to be conserved in other eukaryotes and may be relevant to understanding its role in human disease.