Gametes are highly differentiated cells specialized to carry and protect the parental genetic information. Their chromatin organization is highly compacted and the establishment of this nuclear structure involves many chromatin processes. Technical difficulties exclude the analysis of precise histone mutations during mouse spermatogenesis. The model organism Saccharomyces cerevisiae possesses a differentiation pathway named sporulation with striking similarities with mammalian spermatogenesis. This study took advantage of this yeast pathway and performed a systematic mutational screen on the histone H2A and H2B, revealing the residues which are essential for the formation of spores. Many of them are localized on the lateral side of the nucleosome, highlighting the importance of this surface for meiotic divisions and the formation of spores. Second, histones have been purified at different stages of sporulation and their post-translational modifications analyzed by mass spectrometry. Fifteen new sites of acetylation, methylation or phosphorylation have been identified on the core histones in yeast. Methylation of H2BK37 appears to be a new meiotic mark and is important for Rad51 action. Thus, this modification is conserved during mammalian spermatogenesis when it is found enriched during meiosis. Altogether, our results demonstrate that a combination of genetic and proteomic approaches applied to yeast sporulation can reveal new aspects of chromatin signaling pathways during mammalian spermatogenesis.