Phosphorylation is a key post-translational modification regulating protein function and biological outcomes. However, the phosphorylation dynamics orchestrating mammalian oocyte development remains poorly understood. In the present study, we apply high-resolution mass spectrometry-based phosphoproteomics to obtain the first global in vivo quantification of mouse oocyte phosphorylation. Of more than 8000 phosphosites, 75% significantly oscillate and 64% exhibit marked upregulation during meiotic maturation, indicative of the dominant regulatory role. Moreover, we identify numerous novel phosphosites on oocyte proteins and a few highly conserved phosphosites in oocytes from different species. Through functional perturbations, we demonstrate that phosphorylation status of specific sites participates in modulating critical events including metabolism, translation, and RNA processing during meiosis. Finally, we combine inhibitor screening and enzyme-substrate network prediction to discover previously unexplored kinases and phosphatases that are essential for oocyte maturation. In sum, our data define landscape of the oocyte phosphoproteome, enabling in-depth mechanistic insights into developmental control of germ cells.

中文翻译：

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减数分裂成熟过程中小鼠卵母细胞的整体磷酸化景观。


磷酸化是调节蛋白质功能和生物学结局的关键翻译后修饰。然而，协调哺乳动物卵母细胞发育的磷酸化动力学仍然知之甚少。在本研究中，我们应用基于高分辨率质谱的磷酸化蛋白质组学来获得小鼠卵母细胞磷酸化的首次体内定量。在 8000 多个磷酸化位点中，75% 显著振荡，64% 在减数分裂成熟过程中表现出显著上调，表明具有主导的调节作用。此外，我们在卵母细胞蛋白上鉴定了许多新的磷酸位点，在不同物种的卵母细胞中鉴定了一些高度保守的磷酸位点。通过功能扰动，我们证明特定位点的磷酸化状态参与调节减数分裂过程中的关键事件，包括代谢、翻译和 RNA 加工。最后，我们将抑制剂筛选和酶底物网络预测相结合，以发现以前未探索的对卵母细胞成熟至关重要的激酶和磷酸酶。总之，我们的数据定义了卵母细胞磷酸化蛋白质组的景观，从而能够深入了解生殖细胞的发育控制。