Research Institute for Microbial Diseases, Osaka University, Osaka, Japan.
Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan.
PLoS Genet. 2021 May 7;17(5):e1009412. doi: 10.1371/journal.pgen.1009412. eCollection 2021 May.
Meiosis is a cell division process with complex chromosome events where various molecules must work in tandem. To find meiosis-related genes, we screened evolutionarily conserved and reproductive tract-enriched genes using the CRISPR/Cas9 system and identified potassium channel tetramerization domain containing 19 (Kctd19) as an essential factor for meiosis. In prophase I, Kctd19 deficiency did not affect synapsis or the DNA damage response, and chiasma structures were also observed in metaphase I spermatocytes of Kctd19 KO mice. However, spermatocytes underwent apoptotic elimination during the metaphase-anaphase transition. We were able to rescue the Kctd19 KO phenotype with an epitope-tagged Kctd19 transgene. By immunoprecipitation-mass spectrometry, we confirmed the association of KCTD19 with zinc finger protein 541 (ZFP541) and histone deacetylase 1 (HDAC1). Phenotyping of Zfp541 KO spermatocytes demonstrated XY chromosome asynapsis and recurrent DNA damage in the late pachytene stage, leading to apoptosis. In summary, our study reveals that KCTD19 associates with ZFP541 and HDAC1, and that both KCTD19 and ZFP541 are essential for meiosis in male mice.
减数分裂是一个具有复杂染色体事件的细胞分裂过程，其中各种分子必须协同工作。为了找到与减数分裂相关的基因，我们使用 CRISPR/Cas9 系统筛选了进化上保守和生殖道富集的基因，并鉴定出钾通道四聚化结构域包含 19 个（Kctd19）是减数分裂所必需的一个关键因子。在前期 I 中，Kctd19 缺陷不影响联会或 DNA 损伤反应，中期 I 精母细胞中的 Kctd19 KO 小鼠也观察到交叉结构。然而，精母细胞在中期-后期过渡期间经历凋亡消除。我们能够通过标记表位的 Kctd19 转基因来挽救 Kctd19 KO 表型。通过免疫沉淀-质谱分析，我们证实了 KCTD19 与锌指蛋白 541（ZFP541）和组蛋白去乙酰化酶 1（HDAC1）的关联。Zfp541 KO 精母细胞的表型分析表明，X 染色体和 Y 染色体的联会失败，在晚期粗线期出现反复的 DNA 损伤，导致凋亡。总之，我们的研究揭示了 KCTD19 与 ZFP541 和 HDAC1 相关，并且 KCTD19 和 ZFP541 都是雄性小鼠减数分裂所必需的。
