Global phosphoproteomic analysis identified key kinases regulating male meiosis in mouse
Global phosphoproteomic analysis identified key kinases regulating male meiosis in mouse
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DOI：
10.1007/s00018-022-04507-8
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发表时间：
2022-08-01
影响因子：
8
通讯作者：
Guo, Xuejiang
中科院分区：
文献类型：
--
作者：
Li, Haojie;Chen, Hong;Guo, Xuejiang
减数分裂是从真菌到哺乳动物的一个高度保守的过程，受到蛋白质磷酸化的调控。由于磷酸化丰度低，因此缺乏哺乳动物减数分裂磷酸化调控的系统表征。使用磷酸化蛋白质组学方法，我们描绘了大规模的磷酸化蛋白质组的纯化初级精母细胞减数分裂I，并确定了14,660磷酸化位点的4419磷蛋白。激酶-底物磷酸化网络分析以及体外减数分裂研究表明，CDK 9是减数分裂向中期I进展所必需的，并且在参与减数分裂细胞周期的蛋白质中具有丰富的底物磷酸化位点。此外，组蛋白和表观遗传因子被发现被广泛磷酸化。其中，HASPIN被发现对男性生育力至关重要。Haspin基因敲除可导致染色体错位、中期精母细胞凋亡和精子数量减少，这是由于H3 T3 ph、染色体过客复合体（CPC）和纺锤体组装检查点（SAC）的失调所致。复杂的蛋白磷酸化及其在减数分裂中的重要调控功能表明，深入研究磷酸化介导的信号转导有助于阐明减数分裂的机制。
Meiosis, a highly conserved process in organisms from fungi to mammals, is subjected to protein phosphorylation regulation. Due to the low abundance of phosphorylation, there is a lack of systemic characterization of phosphorylation regulation of meiosis in mammals. Using the phosphoproteomic approach, we profiled large-scale phosphoproteome of purified primary spermatocytes undergoing meiosis I, and identified 14,660 phosphorylation sites in 4419 phosphoproteins. Kinase-substrate phosphorylation network analysis followed by in vitro meiosis study showed that CDK9 was essential for meiosis progression to metaphase I and had enriched substrate phosphorylation sites in proteins involved in meiotic cell cycle. In addition, histones and epigenetic factors were found to be widely phosphorylated. Among those, HASPIN was found to be essential for male fertility. Haspin knockout led to misalignment of chromosomes, apoptosis of metaphase spermatocytes and a decreased number of sperm by deregulation of H3T3ph, chromosomal passenger complex (CPC) and spindle assembly checkpoint (SAC). The complicated protein phosphorylation and its important regulatory functions in meiosis indicated that in-depth studies of phosphorylation-mediated signaling could help us elucidate the mechanisms of meiosis.