Department of Molecular Biology and Microbiology, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA.
BMC Genomics. 2012 Aug 1;13:359. doi: 10.1186/1471-2164-13-359.
Dis3 is ribonuclease that acts directly in the processing, turnover, and surveillance of a large number of distinct RNA species. Evolutionarily conserved from eubacteria to eukaryotes and a crucial component of the RNA processing exosome, Dis3 has been shown to be essential in yeast and fly S2 cells. However, it is not known whether Dis3 has essential functions in a metazoan. This study inquires whether Dis3 is required for Drosophila development and viability and how Dis3 regulates the transcriptome in the developing fly.
Using transgenic flies, we show that Dis3 knock down (Dis3KD) retards growth, induces melanotic tumor formation, and ultimately results in 2nd instar larval lethality. In order to determine whether Dis3KD fly phenotypes were a consequence of disrupting developmentally regulated RNA turnover, we performed RNA deep sequencing analysis on total RNA isolated from developmentally staged animals. Bioinformatic analysis of transcripts from Dis3KD flies reveals substantial transcriptomic changes, most notably down-regulation in early expressed RNAs. Finally, gene ontology analysis of this early stage shows that Dis3 regulates transcripts related to extracellular structure and remodelling, neurogenesis, and nucleotide metabolism.
We conclude that Dis3 is essential for early Drosophila melanogaster development and has specific and important stage-specific roles in regulating RNA metabolism. In showing for the first time that Dis3 is required for the development of a multicellular organism, our work provides mechanistic insight into how Dis3-either independent of or associated with the RNA processing exosome-participates in cell type-specific RNA turnover in metazoan development.
Dis3 是一种核糖核酸酶，可直接作用于大量不同 RNA 物种的加工、周转和监测。从细菌到真核生物，Dis3 在进化上是保守的，并且是 RNA 加工外切体的关键组成部分，已被证明在酵母和果蝇 S2 细胞中是必不可少的。然而，尚不清楚 Dis3 在后生动物中是否具有必需的功能。本研究探讨了 Dis3 是否是果蝇发育和生存所必需的，以及 Dis3 如何调节发育过程中的转录组。
利用转基因果蝇，我们发现 Dis3 敲低（Dis3KD）会减缓生长速度、诱导黑色素瘤形成，并最终导致 2 龄幼虫致死。为了确定 Dis3KD 果蝇表型是否是破坏发育调控的 RNA 周转的结果，我们对从发育阶段的动物中分离的总 RNA 进行了 RNA 深度测序分析。Dis3KD 果蝇转录本的生物信息学分析显示转录组发生了大量变化，尤其是早期表达的 RNA 下调。最后，对早期阶段的基因本体分析表明，Dis3 调节与细胞外结构和重塑、神经发生和核苷酸代谢相关的转录物。
我们得出结论，Dis3 对早期果蝇发育是必需的，并且在调节 RNA 代谢方面具有特定的和重要的阶段特异性作用。本研究首次表明 Dis3 是多细胞生物发育所必需的，为 Dis3 参与后生动物发育中特定细胞类型的 RNA 周转的机制提供了深入的了解，这种作用可以独立于或与 RNA 加工外切体相关。
