1. KCTD19 and its associated protein ZFP541 are independently essential for meiosis in male mice
- Author
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Chisato Kodera, Kei-ichiro Ishiguro, Takayuki Koyano, Masahito Ikawa, Makoto Matsuyama, Seiya Oura, and Yuki Horisawa-Takada
- Subjects
Male ,Cancer Research ,Cell division ,Chromosomal Proteins, Non-Histone ,Cell Cycle Proteins ,Histone Deacetylase 1 ,Apoptosis ,Immunostaining ,QH426-470 ,Mice ,0302 clinical medicine ,Spermatocytes ,Animal Cells ,Testis ,Medicine and Health Sciences ,Transgenes ,Cell Cycle and Cell Division ,Meiotic Prophase I ,Genetics (clinical) ,Conserved Sequence ,Zinc finger ,Staining ,0303 health sciences ,Genes, Essential ,Cell Death ,Chromosome Biology ,Genetically Modified Organisms ,Synapsis ,Nuclear Proteins ,Seminiferous Tubules ,Spermatids ,Chiasma ,Cell biology ,Precipitation Techniques ,Meiosis ,Phenotype ,Cell Processes ,Engineering and Technology ,Cellular Types ,Anatomy ,Genetic Engineering ,Genital Anatomy ,Research Article ,Biotechnology ,DNA damage ,Bioengineering ,Biology ,Research and Analysis Methods ,Evolution, Molecular ,03 medical and health sciences ,Genetics ,Animals ,Immunoprecipitation ,Molecular Biology ,Metaphase ,Ecology, Evolution, Behavior and Systematics ,030304 developmental biology ,Cell Nucleus ,Genetically Modified Animals ,Reproductive System ,Chromosome ,Biology and Life Sciences ,Cell Biology ,Sperm ,Chromosome Pairing ,Fertility ,Germ Cells ,Specimen Preparation and Treatment ,Pachytene Stage ,CRISPR-Cas Systems ,Anaphase ,030217 neurology & neurosurgery ,DNA Damage ,Transcription Factors - 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., Author summary Meiosis is a fundamental process that consists of one round of genomic DNA replication and two rounds of chromosome segregation, producing four haploid cells. To properly distribute their genetic material, cells need to undergo complex chromosome events such as a physical linkage of homologous chromosomes (termed synapsis) and meiotic recombination. The molecules involved in these events have not been fully characterized yet, especially in mammals. 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. Further, we confirmed the association of KCTD19 with zinc finger protein 541 (ZFP541) and histone deacetylase 1 (HDAC1). By observing meiosis of Zfp541 knockout germ cells, we found that Zfp541 was also essential for meiosis. These results show that the KCTD19/ZFP541 complex plays a critical role and is indispensable for male meiosis and fertility.
- Published
- 2021