通过Pumilio和CCR4-NOT死亡酶复合物的Drosophila转录组的调节
Rebecca J Haugen1, Catherine Barnier2, Nathan D Elrod3
1Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota 55455, USA.
概括
这项研究揭示了Drosophila发育中的Pumilio (Pum) 蛋白调节的广泛的信使RNA (mRNA) 网络. 它确定了新的直接Pum目标,并阐明了Pum介导基因调节的机制.
科学领域:
- 发展生物学 发展生物学
- 分子生物学分子生物学
- 遗传学 遗传学 是一个
背景情况:
- 序列特异性的RNA结合蛋白Pumilio (Pum) 对于Drosophila发育至关重要.
- 由Pum调节的mRNAs的完整网络在很大程度上仍然没有被描述.
研究的目的:
- 为了全面识别直接的Pum目标基因,并了解Drosophila中的Pum的调节机制.
- 调查CCR4-NOT死亡酶复合体在Pum介导抑制中的作用.
主要方法:
- 利用敲除和敲除方法与RNA测序 (RNA-seq) 结合,以评估Pum对转录组的影响.
- 采用了改进的RNA共免疫沉方法来识别Drosophila胚胎中的Pum结合的mRNA.
- 集成的转录基因数据与Pum结合基因的位置,用于新型目标基因的识别.
主要成果:
- 确定了参与神经,肌肉,翅膀,生殖细胞发育和细胞增殖的新型直接Pum点基因,包括Wnt,TGF-β,MAPK/ERK和Notch信号通路的组成部分.
- 发现了Pum和CCR4-NOT死亡酶复合体所准的mRNAs的一致调节.
- 计算建模表明Pum结合特征 (站点号,集群,上下文) 是关键的监管决定因素,而不是代码子的最佳性.
结论:
- 这项工作阐明了Pum广泛的调节网络及其对Drosophila多样化的发育过程的影响.
- 这些发现为控制Pum介导基因调节有效性的参数提供了新的见解,挑战了关于CCR4-NOT在mRNA降解中的作用的先前模型.
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