Male germ cells experience a drastic chromatin remodeling through the nucleo-histone to nucleo-protamine (NH-NP) transition necessary for proper sperm functionality. Post-translational modifications (PTMs) of H4 Lys5, such as acetylation (H4K5ac), play a crucial role in epigenetic control of nucleosome disassembly facilitating protamine incorporation into paternal DNA. It has been shown that butyrylation on the same residue (H4K5bu) participates in temporal regulation of NH-NP transition in mice, delaying the bromodomain testis specific protein (BRDT)-dependent nucleosome disassembly and potentially marking retained nucleosomes. However, no information was available so far on this modification in human sperm. Here, we report a dual behavior of H4K5bu and H4K5ac in human normal spermatogenesis, suggesting a specific role of H4K5bu during spermatid elongation, coexisting with H4K5ac although with different starting points. This pattern is stable under different testicular pathologies, suggesting a highly conserved function of these modifications. Despite a drastic decrease of both PTMs in condensed spermatids, they are retained in ejaculated sperm, with 30% of non-colocalizing nucleosome clusters, which could reflect differential paternal genome retention. Whereas no apparent effect of these PTMs was observed associated with sperm quality, their presence in mature sperm could entail a potential role in the zygote.

Biochemistry and Molecular Genetics Service Clínic Barcelona 08036 Barcelona Spain

Clinic Institute of Gynaecology Obstetrics and Neonatology Clínic Barcelona 08036 Barcelona Spain

Clinic Institute of Urology and Nephrology IDIBAPS Clínic Barcelona 08036 Barcelona Spain

Department of Pathology Clínic Barcelona 08036 Barcelona Spain

Department of Zoology Faculty of Science Charles University 128 44 Prague Czech Republic

Faculty of Medicine University of Vic Central University of Catalonia 08500 Barcelona Spain

Laboratory of Reproductive Biology Institute of Biotechnology Czech Academy of Sciences BIOCEV 252 50 Vestec Czech Republic

Molecular Biology of Reproduction and Development Research Group Institut d'Investigacions Biomèdiques August Pi i Sunyer 08036 Barcelona Spain

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