CPEB and miR-15/16 Co-Regulate Translation of Cyclin E1 mRNA during Xenopus Oocyte Maturation.
CPEB and miR-15/16 Co-Regulate Translation of Cyclin E1 mRNA during Xenopus Oocyte Maturation.
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DOI：
10.1371/journal.pone.0146792
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Standart N
中科院分区：
文献类型：
--
作者：
Wilczynska A;Git A;Argasinska J;Belloc E;Standart N
非洲爪蟾卵母细胞减数分裂成熟以及早期胚胎发生过程中的细胞周期转换在储存的无活性母源mRNA水平上受到严格调控。在此我们研究了细胞周期蛋白E1的翻译调控，它对于未受精卵的中期II阻滞以及早期胚胎中S期的启动是必需的。我们表明细胞周期蛋白E1的mRNA受到细胞质多聚腺苷酸化元件（CPEs）以及其3’非翻译区（3’UTR）内的两个miR - 15/16靶位点的调控。此外，我们提供的证据表明母源miR - 15/16微小RNA与CPE结合蛋白（CPEB）共同免疫沉淀，并且CPEB与RNA诱导沉默复合体（RISC）成分Ago2相互作用。使用竞争RNA和突变的细胞周期蛋白E1的3’UTR进行的实验表明，在成熟的早期阶段，当CPEB变得有限且受抑制的mRNA的细胞质多聚腺苷酸化开始时，调控元件协同作用以维持对细胞周期蛋白E1的mRNA的抑制。重要的是，注射抗miR - 15/16锁核酸（LNA）会导致减数分裂成熟过程中内源性细胞周期蛋白E1的mRNA早期多聚腺苷酸化，并加速生发泡破裂（GVBD），这一切都有力地表明近端CPEB和微小RNA - 蛋白复合体（miRNP）相互作用以稳定彼此。我们得出结论，miR - 15/16和CPEB共同调控细胞周期蛋白E1的mRNA。这是对这两种途径协同作用的首次证明。
Cell cycle transitions spanning meiotic maturation of the Xenopus oocyte and early embryogenesis are tightly regulated at the level of stored inactive maternal mRNA. We investigated here the translational control of cyclin E1, required for metaphase II arrest of the unfertilised egg and the initiation of S phase in the early embryo. We show that the cyclin E1 mRNA is regulated by both cytoplasmic polyadenylation elements (CPEs) and two miR-15/16 target sites within its 3’UTR. Moreover, we provide evidence that maternal miR-15/16 microRNAs co-immunoprecipitate with CPE-binding protein (CPEB), and that CPEB interacts with the RISC component Ago2. Experiments using competitor RNA and mutated cyclin E1 3’UTRs suggest cooperation of the regulatory elements to sustain repression of the cyclin E1 mRNA during early stages of maturation when CPEB becomes limiting and cytoplasmic polyadenylation of repressed mRNAs begins. Importantly, injection of anti-miR-15/16 LNA results in the early polyadenylation of endogenous cyclin E1 mRNA during meiotic maturation, and an acceleration of GVBD, altogether strongly suggesting that the proximal CPEB and miRNP complexes act to mutually stabilise each other. We conclude that miR-15/16 and CPEB co-regulate cyclin E1 mRNA. This is the first demonstration of the co-operation of these two pathways.