ZFP541 is indispensable for pachytene progression by interacting with KCTD19 and activates meiotic gene expression in mouse spermatogenesis

Meiosis is essential for fertility in sexually reproducing species, extensive studies tried to delineate this sophisticated process. Notwithstanding, the molecules involved in meiosis have not been fully characterized. In this study, we investigate the role of zinc finger protein 541 (ZFP541) and its interacting protein potassium channel tetramerization domain containing 19 (KCTD19) in mice. We demonstrate that they are indispensable for male fertility by regulating proper pachytene progression. ZFP541 is expressed starting from leptotene to round spermatids, and KCTD19 is initially expressed in pachytene. Depletion of Zfp541 or Kctd19 leads to infertility in male mice, and exhibits retarded progression from early to mid/late pachynema. In addition, Zfp541-/- spermatocytes show abnormal programmed DNA double-strand breaks (DSBs) repair, impaired crossover formation/resolution, and asynapsis of the XY chromosomes. Immunoprecipitation-mass spectrometry (IP-MS) and in vitro Co-IP reveal that ZFP541 interacts with KCTD19, histone deacetylase 1/2 (HDAC1), HDAC2 and deoxynucleotidyltransferase terminal-interacting protein 1 (DNTTIP1). Furthermore, RNA-seq and CUT&Tag analyses demonstrate that ZFP541 binds to the promoter regions of genes involved in meiosis and post-meiosis including Kctd19, and activates their transcription. Taken together, our studies reveal a ZFP541-Kctd19 transcription regulatory axis and the crucial role of ZFP541 and KCTD19 for pachytene progression and fertility in male mice.