NAT10-mediated N4-Acetylcytidine is Required for Spermatogenesis and Male Fertility

RNA modifications are rapidly emerging as critical post-transcriptional regulators which play a key role in various biological processes. N4-acetylcysteine (ac4C) is one of the “epi-transcriptome” that is present on tRNA, rRNA, mRNA and is also highly conserved in all species. However, cytidine acetylation's in vivo physiological functions remain poorly understood particularly in mammals. Here we demonstrate only known ac4C “writer”—N-acetyltransferase 10 (NAT10) plays essential roles in male reproduction. NAT10 protein and the ac4C modification have different patterns in different organs and there are dynamic changes during spermatogenesis. Germ cell-specific ablation of NAT10 severely inhibited meiotic entry and defects in synapse the homologous chromosomes, repair of DNA double-strand breaks (DSBs) during prophase of the first meiotic division. ac4C RIP-qPCR reveals genes functioning in meiotic prophase I show a decrease in Nat10fl/-; Stra8 GFP-Cre mouse testes. These findings highlight the crucial physiological functions of mRNA ac4C modification in male spermatogenesis and expand our understanding of mRNA ac4C modification in the regulation of specific physiological processes in vivo. Overall design: Examination of WT and Nat10-SKO transcriptome: A total of 17 samples were analyzed in this study, including WT and Nat10-SKO spermatogenic cells from 3 stages, spermatogonia, preleptotene and leptotene/zygotene stage.