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Mobilization of Dormant Cnot7 mRNA Promotes Deadenylation of Maternal Transcripts During Mouse Oocyte Maturation

机译:在小鼠卵母细胞成熟过程中休眠Cnot7 mRNA的动员促进母本转录产物的腺苷酸化。

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摘要

Maternal mRNAs in oocytes are remarkably stable. In mouse, oocyte maturation triggers a transition from mRNA stability to instability. This transition is a critical event in the oocyte-to-embryo transition in which a differentiated oocyte loses its identity as it is transformed into totipotent blastomeres. We previously demonstrated that phosphorylation of MSY2, an RNA-binding protein, and mobilization of mRNAs encoding the DCP1A-DCP2 decapping complex contribute to maternal mRNA destruction during meiotic maturation. We report here that Cnot7, Cnot6l, and Pan2, key components of deadenylation machinery, are also dormant maternal mRNAs that are recruited during oocyte maturation. Inhibiting the maturation-associated increase in CNOT7 (or CNOT6L) using a small interference RNA approach inhibits mRNA deadenylation, whereas inhibiting the increase in PAN2 has little effect. Reciprocally, expressing CNOT7 (or CNOT6L) in oocytes prevented from resuming meiosis initiates deadenylation of mRNAs. These effects on deadenylation are also observed when the total amount of poly (A) is quantified. Last, inhibiting the increase in CNOT7 protein results in an ∼70% decrease in transcription in 2-cell embryos.
机译:卵母细胞中的母体mRNA非常稳定。在小鼠中,卵母细胞成熟会触发从mRNA稳定性到不稳定的转变。该转变是卵母细胞向胚胎转变的关键事件,在该过程中,分化的卵母细胞由于转化为全能卵裂球而失去其身份。我们以前证明MSY2,一种RNA结合蛋白的磷酸化和编码DCP1A-DCP2脱盖复合物的mRNA的动员有助于减数分裂成熟期间母体mRNA的破坏。我们在这里报告,Cnot7,Cnot6l和Pan2,去甲基化机制的关键组成部分,也是在卵母细胞成熟过程中募集的休眠母体mRNA。使用小的干扰RNA方法抑制CNOT7(或CNOT6L)与成熟相关的增加会抑制mRNA的腺苷酸化,而抑制PAN2的增加几乎没有效果。相反地​​,在被阻止恢复减数分裂的卵母细胞中表达CNOT7(或CNOT6L)会引发mRNA的去甲腺苷酸化。当对聚(A)的总量进行定量时,也观察到了对腺苷酸化的这些影响。最后,抑制CNOT7蛋白的增加会导致2细胞胚胎转录降低约70%。

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