2P-10 PDF
MS-based characterization of post-translational modifications on H3 variants in mouse testes
Spermatogenesis is a biological process in which sperm are produced in the testes. Various histone variants, including testis-specific histones, are detected and expressed during this stage. Post-translational modifications (PTMs) are decorated on histone variants and they have been reported to be associated with chromatin remodeling during spermatogenesis. Although some PTMs on H3 variants during spermatogenesis have been detected by proteomic approaches, variant-specific PTMs are still underexplored. In this research, we comprehensively characterized PTMs on histone H3 variants by MS-based approaches in mouse testes. Histone H3 variants were separated using HPLC, utilizing a reversed-phase column with ion-pairing reagent. H3 variants and their PTMs were characterized by top-down analysis using MALDI-TOF/MS and bottom-up strategy using nano-LC-MS/MS. PTMs, including methylation and acetylation, on H3 variants in mouse testes were identified. In particular, various lysine modifications on testis-specific H3 histone (H3t) were detected by our approach. Moreover, we detected formylation as a novel PTM on H3 variants in mouse testes. Our data will provide information for future studies, which can be applied for identifying other samples to detect novel or specific patterns.