Department of Chromosome Biology, Institute of Molecular Embryology and Genetics (IMEG), Kumamoto University, Kumamoto, 860-0811, Japan.
Division of Gene Regulation, Institute for Advanced Medical Research, Keio University School of Medicine, 160-8582, Tokyo, Japan.
Sci Rep. 2020 Jun 22;10(1):10094. doi: 10.1038/s41598-020-67116-0.
FZR1/CDH1 is an activator of Anaphase promoting complex/Cyclosome (APC/C), best known for its role as E3 ubiquitin ligase that drives the cell cycle. APC/C activity is regulated by CDK-mediated phosphorylation of FZR1 during mitotic cell cycle. Although the critical role of FZR1 phosphorylation has been shown mainly in yeast and in vitro cell culture studies, its biological significance in mammalian tissues in vivo remained elusive. Here, we examined the in vivo role of FZR1 phosphorylation using a mouse model, in which non-phosphorylatable substitutions were introduced in the putative CDK-phosphorylation sites of FZR1. Although ablation of FZR1 phosphorylation did not show substantial consequences in mouse somatic tissues, it led to severe testicular defects resulting in male infertility. In the absence of FZR1 phosphorylation, male juvenile germ cells entered meiosis normally but failed to enter meiosis II or form differentiated spermatids. In aged testis, male mutant germ cells were overall abolished, showing Sertoli cell-only phenotype. In contrast, female mutants showed apparently normal progression of meiosis. The present study demonstrated that phosphorylation of FZR1 is required for temporal regulation of APC/C activity at meiosis II entry, and for maintenance of spermatogonia, which raised an insight into the sexual dimorphism of FZR1-regulation in germ cells.
FZR1/CDH1 是后期促进复合物/细胞周期蛋白体(APC/C)的激活剂,以作为 E3 泛素连接酶驱动细胞周期的作用而闻名。APC/C 的活性通过有丝分裂细胞周期中 CDK 介导的 FZR1 磷酸化来调节。虽然 FZR1 磷酸化的关键作用主要在酵母和体外细胞培养研究中得到了证实,但在哺乳动物组织体内的生物学意义仍然难以捉摸。在这里,我们使用小鼠模型研究了 FZR1 磷酸化的体内作用,其中在 FZR1 的假定 CDK 磷酸化位点引入了不可磷酸化的取代。尽管 FZR1 磷酸化的缺失在小鼠体细胞组织中没有显示出实质性的后果,但它导致了严重的睾丸缺陷,导致男性不育。在缺乏 FZR1 磷酸化的情况下,雄性幼体生殖细胞正常进入减数分裂,但无法进入减数分裂 II 或形成分化的精母细胞。在老年睾丸中,雄性突变体生殖细胞总体上被消除,表现出唯支持细胞表型。相比之下,雌性突变体显示出减数分裂的正常进展。本研究表明,FZR1 的磷酸化对于 APC/C 活性在减数分裂 II 进入时的时间调节以及精原细胞的维持是必需的,这揭示了生殖细胞中 FZR1 调节的性别二态性的一个新见解。