H4K5 Butyrylation Coexist with Acetylation during Human Spermiogenesis and Are Retained in the Mature Sperm Chromatin

Alberto de la Iglesia; Paula Jauregi; Meritxell Jodar; Ferran Barrachina; Lukas Ded; Carme Mallofré; Leonardo Rodríguez-Carunchio; Juan Manuel Corral; Josep Lluís Ballescà; Katerina Komrskova; Judit Castillo; Rafael Oliva

doi:10.3390/ijms232012398

H4K5 Butyrylation Coexist with Acetylation during Human Spermiogenesis and Are Retained in the Mature Sperm Chromatin

Int J Mol Sci. 2022 Oct 17;23(20):12398. doi: 10.3390/ijms232012398.

Authors

Affiliations

¹ Molecular Biology of Reproduction and Development Research Group, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Fundació Clínic per a la Recerca Biomèdica, Departament de Biomedicina, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona (UB), 08036 Barcelona, Spain.
² Biochemistry and Molecular Genetics Service, Clínic Barcelona, 08036 Barcelona, Spain.
³ Laboratory of Reproductive Biology, Institute of Biotechnology, Czech Academy of Sciences, BIOCEV, 252 50 Vestec, Czech Republic.
⁴ Department of Pathology, Clínic Barcelona, 08036 Barcelona, Spain.
⁵ Faculty of Medicine, University of Vic-Central University of Catalonia (UVic-UCC), 08500 Barcelona, Spain.
⁶ Clinic Institute of Urology and Nephrology, IDIBAPS, Clínic Barcelona, 08036 Barcelona, Spain.
⁷ Clinic Institute of Gynaecology, Obstetrics and Neonatology, Clínic Barcelona, 08036 Barcelona, Spain.
⁸ Department of Zoology, Faculty of Science, Charles University, 128 44 Prague, Czech Republic.

Abstract

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.

Keywords: H4K5; acetylation; butyrylation; epigenetic regulation; sperm; sperm chromatin; spermatogenesis.

MeSH terms

Acetylation
Animals
Chromatin Assembly and Disassembly
Chromatin* / metabolism
Histones / metabolism
Humans
Male
Mice
Nucleosomes* / metabolism
Protamines / metabolism
Protein Processing, Post-Translational
Semen / metabolism
Spermatids / metabolism
Spermatogenesis / physiology
Spermatozoa / metabolism

Substances

Chromatin
Nucleosomes
Histones
Protamines

Grants and funding

This research was funded by grants PI16/00346 and PI20/00936 to R.O. from the ‘Ministerio de Economía y Competividad’ (Spain, ‘fondos FEDER, una manera de hacer Europa’). This publication is also based upon work from COST Action CA20119 (ANDRONET) supported by COST ((European Cooperation in Science and Technology) www.cost.eu, accessed on 6 September 2022). J.C. was supported by the Sara Borrell Postdoctoral Fellowship, ‘Acción Estratégica en Salud’, CD17/00109. J.C. is a Serra Húnter fellow (Universitat de Barcelona, Generalitat de Catalunya). A.I. is supported by a fellowship from the ‘Ministerio de Economía, Industria y Competitividad’ (Spain), ‘Acción Estratégica en Salud, contrato Predoctoral de Formación en Investigación en Salud’ (PFIS, FI17/00224). F.B. received a grant from the Ministerio de Educación, Cultura y Deporte para la Formación de Profesorado Universitario (Spain) (FPU15/02306). M.J. was supported by the Government of Catalonia (Generalitat de Catalunya, pla estratègic de recerca i innovació en salut, PERIS 2016-2020, SLT002/16/00337). L.D. and K.K. were supported by Czech Science Foundation (GACR GJ20-17403Y) to L.D. and GC20-20217J to K.K., project BIOCEV (CZ.1.05/1.1.00/02/0109) from the ERDF and the Institute of Biotechnology RVO:86652036 to L.D. and K.K.