KCTD19 and its associated protein ZFP541 are independently essential for meiosis in male mice

Seiya Oura,Kei-Ichiro Ishiguro,Masahito Ikawa,Makoto Matsuyama,Yuki Horisawa-Takada,Chisato Kodera,Takayuki Koyano,Paula E Cohen

doi:10.1371/journal.pgen.1009412

Abstract

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.

Highlights

Meiosis is a division process consisting of one round of DNA replication and two rounds of chromosome segregation, producing four haploid gametes
Using a CRISPR/Cas9-screening system, we identified the potassium channel tetramerization domain containing 19 (Kctd19) as an essential factor for meiosis in male mice
To investigate the spatial expression of Kctd19 in mice, we performed multi-tissue RT-Polymerase chain reaction (PCR) using cDNA obtained from adult tissues and embryonic ovary, and we found that Kctd19 was

Summary

Introduction

Meiosis is a division process consisting of one round of DNA replication and two rounds of chromosome segregation, producing four haploid gametes. The homologs begin to pair and synapse, followed by meiotic recombination yielding a physical tether between homologs (chiasmata) After completing these chromosome events, the cells transition to the first meiotic division, where homologs are segregated to the opposite poles, followed by the segregation of sister chromatids in the round of cell division. We have generated over 300 testis-enriched gene KO mice with conventional ES cell-mediated and the CRISPR/Cas9-mediated methods [2,3,4,5] and showed about one-third of them are indispensable for male fertility [6,7,8] During this phenotypic screening, we identified potassium channel tetramerization domain containing 19 (Kctd19) as an evolutionarily conserved and testis expressed gene that is essential for male fertility in mice

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Discussion

Conclusion