Key Laboratory for Stem Cells and Tissue Engineering (Sun Yat-sen University), Ministry of Education, Guangzhou, China.
Department of histology and embryology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China.
J Cell Physiol. 2023 Jul;238(7):1580-1591. doi: 10.1002/jcp.31037. Epub 2023 Jun 12.
Poly(ADP-ribosyl)ation (PARylation) is an important post-translational modification of proteins that involves the transfer of ADP-ribose moieties, and plays important roles in many biological processes including DNA repair, gene expression, RNA processing, ribosome biogenesis, and protein translation. Though it is accepted that PARylation is crucial for oocyte maturation, little is known about how Mono(ADP-ribosyl)ation (MARylation) regulates this process. Here, we report that Parp12, a mon(ADP-ribosyl) transferase of poly(ADP-ribosyl) polymerase (PARP) family, was highly expressed at all stages of oocytes during meiotic maturation. At germinal vesicle (GV) stage, PARP12 was mainly distributed in cytoplasm. Interestingly, PARP12 formed granular aggregation near to spindle poles during metaphase I (MI) and metaphase II (MII). PARP12 depletion results in abnormal spindle organization and chromosome misalignment in mouse oocytes. Chromosome aneuploidy frequency in PARP12 knockdown oocytes was significantly increased. Importantly, PARP12 knockdown triggers activation of spindle assembly checkpoint as shown by active BUBR1 in PARP12-KD MI oocytes. Besides, F actin was significantly attenuated in PARP12-KD MI oocytes which may affect the asymmetric division process. Transcriptomic analysis demonstrated that PARP12 depletion disrupts transcriptome homeostasis. Collectively, our results showed that the maternally expressed mono(ADPribosyl) transferases PARP12 was essential for oocyte meiotic maturation in mouse.
多聚(ADP-核糖)化(PARylation)是一种重要的蛋白质翻译后修饰,涉及 ADP-核糖基的转移,在许多生物过程中发挥重要作用,包括 DNA 修复、基因表达、RNA 加工、核糖体生物发生和蛋白质翻译。尽管人们普遍认为 PARylation 对卵母细胞成熟至关重要,但对于单聚(ADP-核糖)化(MARylation)如何调节这一过程知之甚少。在这里,我们报告 Parp12,多聚(ADP-核糖)聚合酶(PARP)家族的单聚(ADP-核糖)转移酶,在减数分裂成熟过程中卵母细胞的所有阶段都高度表达。在生发泡(GV)期,PARP12 主要分布在细胞质中。有趣的是,PARP12 在中期 I(MI)和中期 II(MII)期间在纺锤体极附近形成颗粒聚集。PARP12 耗竭导致小鼠卵母细胞纺锤体组织异常和染色体错位。PARP12 敲低卵母细胞中的染色体非整倍体频率显著增加。重要的是,PARP12 敲低触发纺锤体组装检查点的激活,如 PARP12-KD MI 卵母细胞中的活性 BUBR1 所示。此外,PARP12-KD MI 卵母细胞中的 F 肌动蛋白明显减弱,这可能会影响不对称分裂过程。转录组分析表明,PARP12 耗竭破坏了转录组的平衡。总之,我们的结果表明,母源性表达的单聚(ADP-核糖)转移酶 PARP12 对于小鼠卵母细胞减数分裂成熟是必不可少的。