Histone variant H3.5 in testicular cell differentiation and its interactions with histone chaperones

Abstract Testicular cell differentiation is a highly regulated process, essential for male reproductive health. The histone variant H3.5 is apparently a critical player in this intricate orchestra of cell types, but its regulation and function remains poorly understood. To elucidate its role, we fra...

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Main Authors: Patrick Philipp Weil, Anton Pembaur, Beatrice Wirth, Eda Oetjen, Hannes Büsscher, Klemens Zirngibl, Malte Czarnetzki, Stella Braun, Jann-Frederik Cremers, Daniel Gödde, Stephan Degener, Jan Postberg
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Language:English
Published: Nature Portfolio 2024-12-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-024-83206-9
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author Patrick Philipp Weil
Anton Pembaur
Beatrice Wirth
Eda Oetjen
Hannes Büsscher
Klemens Zirngibl
Malte Czarnetzki
Stella Braun
Jann-Frederik Cremers
Daniel Gödde
Stephan Degener
Jan Postberg
author_facet Patrick Philipp Weil
Anton Pembaur
Beatrice Wirth
Eda Oetjen
Hannes Büsscher
Klemens Zirngibl
Malte Czarnetzki
Stella Braun
Jann-Frederik Cremers
Daniel Gödde
Stephan Degener
Jan Postberg
author_sort Patrick Philipp Weil
collection DOAJ
description Abstract Testicular cell differentiation is a highly regulated process, essential for male reproductive health. The histone variant H3.5 is apparently a critical player in this intricate orchestra of cell types, but its regulation and function remains poorly understood. To elucidate its role, we fractionized testicular cells using c-Kit/CD117 as a separation marker and analyzed H3.5 expression. Further, we investigated the regulation of H3.5 expression using public data repositories. We explored DNA methylation patterns in specific regions of the H3-5 gene and assessed H3-5 copy number gain in seminoma specimens. Additionally, we examined the testicular localization of H3.5 and its histone chaperone interactions to understand its regulation at the protein level. We used qRT-PCR, MeDIP, and qPCR to study H3.5 expression and DNA methylation in various cell types. H3-5 copy number gain was analyzed using qPCR. Protein interactions were investigated through fluorescence-2-hybrid assays in baby hamster kidney cells. H3.5 is primarily enriched in spermatocytes. DNA methylation of a CpG island overlapping the H3-5 promoter appeared to be involved in the tissue-specific regulation of H3.5 expression. Elevated H3.5 expression was observed in seminoma specimens, suggesting a potential link to testicular tumors. H3-5 copy number gain was associated with elevated H3.5 expression in seminoma specimens. Furthermore, we identified physical interactions between H3.5 and histone chaperones Asf1a and Asf1b, HIRA, CAF p150 and DAXX, shedding light on the protein-level regulation of H3.5. These findings provide valuable insights into the molecular mechanisms governing testicular cell differentiation and the potential role of H3.5 in testicular pathologies.
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spelling doaj-art-d24338fcb97d449db64ff57d4539c61a2024-12-22T12:27:22ZengNature PortfolioScientific Reports2045-23222024-12-0114111510.1038/s41598-024-83206-9Histone variant H3.5 in testicular cell differentiation and its interactions with histone chaperonesPatrick Philipp Weil0Anton Pembaur1Beatrice Wirth2Eda Oetjen3Hannes Büsscher4Klemens Zirngibl5Malte Czarnetzki6Stella Braun7Jann-Frederik Cremers8Daniel Gödde9Stephan Degener10Jan Postberg11Clinical Molecular Genetics and Epigenetics, Faculty of Health, Centre for Biomedical Education & Research (ZBAF), Witten/Herdecke UniversityClinical Molecular Genetics and Epigenetics, Faculty of Health, Centre for Biomedical Education & Research (ZBAF), Witten/Herdecke UniversityClinical Molecular Genetics and Epigenetics, Faculty of Health, Centre for Biomedical Education & Research (ZBAF), Witten/Herdecke UniversityClinical Molecular Genetics and Epigenetics, Faculty of Health, Centre for Biomedical Education & Research (ZBAF), Witten/Herdecke UniversityClinical Molecular Genetics and Epigenetics, Faculty of Health, Centre for Biomedical Education & Research (ZBAF), Witten/Herdecke UniversityClinical Molecular Genetics and Epigenetics, Faculty of Health, Centre for Biomedical Education & Research (ZBAF), Witten/Herdecke UniversityClinical Molecular Genetics and Epigenetics, Faculty of Health, Centre for Biomedical Education & Research (ZBAF), Witten/Herdecke UniversityClinical Molecular Genetics and Epigenetics, Faculty of Health, Centre for Biomedical Education & Research (ZBAF), Witten/Herdecke UniversityCentre of Reproductive Medicine and Andrology, University Hospital of MünsterChair of Pathology, Centre for Clinical and Translational Research (ZFKM), Helios University Hospital Wuppertal, Witten/Herdecke UniversityChair of Urology, Centre for Clinical and Translational Research (ZFKM), Helios University Hospital Wuppertal, Witten/Herdecke UniversityClinical Molecular Genetics and Epigenetics, Faculty of Health, Centre for Biomedical Education & Research (ZBAF), Witten/Herdecke UniversityAbstract Testicular cell differentiation is a highly regulated process, essential for male reproductive health. The histone variant H3.5 is apparently a critical player in this intricate orchestra of cell types, but its regulation and function remains poorly understood. To elucidate its role, we fractionized testicular cells using c-Kit/CD117 as a separation marker and analyzed H3.5 expression. Further, we investigated the regulation of H3.5 expression using public data repositories. We explored DNA methylation patterns in specific regions of the H3-5 gene and assessed H3-5 copy number gain in seminoma specimens. Additionally, we examined the testicular localization of H3.5 and its histone chaperone interactions to understand its regulation at the protein level. We used qRT-PCR, MeDIP, and qPCR to study H3.5 expression and DNA methylation in various cell types. H3-5 copy number gain was analyzed using qPCR. Protein interactions were investigated through fluorescence-2-hybrid assays in baby hamster kidney cells. H3.5 is primarily enriched in spermatocytes. DNA methylation of a CpG island overlapping the H3-5 promoter appeared to be involved in the tissue-specific regulation of H3.5 expression. Elevated H3.5 expression was observed in seminoma specimens, suggesting a potential link to testicular tumors. H3-5 copy number gain was associated with elevated H3.5 expression in seminoma specimens. Furthermore, we identified physical interactions between H3.5 and histone chaperones Asf1a and Asf1b, HIRA, CAF p150 and DAXX, shedding light on the protein-level regulation of H3.5. These findings provide valuable insights into the molecular mechanisms governing testicular cell differentiation and the potential role of H3.5 in testicular pathologies.https://doi.org/10.1038/s41598-024-83206-9SpermatogenesisHistone variantsEpigenetic regulationHistone chaperonesChromatin remodelingEpigenetic Plasticity
spellingShingle Patrick Philipp Weil
Anton Pembaur
Beatrice Wirth
Eda Oetjen
Hannes Büsscher
Klemens Zirngibl
Malte Czarnetzki
Stella Braun
Jann-Frederik Cremers
Daniel Gödde
Stephan Degener
Jan Postberg
Histone variant H3.5 in testicular cell differentiation and its interactions with histone chaperones
Scientific Reports
Spermatogenesis
Histone variants
Epigenetic regulation
Histone chaperones
Chromatin remodeling
Epigenetic Plasticity
title Histone variant H3.5 in testicular cell differentiation and its interactions with histone chaperones
title_full Histone variant H3.5 in testicular cell differentiation and its interactions with histone chaperones
title_fullStr Histone variant H3.5 in testicular cell differentiation and its interactions with histone chaperones
title_full_unstemmed Histone variant H3.5 in testicular cell differentiation and its interactions with histone chaperones
title_short Histone variant H3.5 in testicular cell differentiation and its interactions with histone chaperones
title_sort histone variant h3 5 in testicular cell differentiation and its interactions with histone chaperones
topic Spermatogenesis
Histone variants
Epigenetic regulation
Histone chaperones
Chromatin remodeling
Epigenetic Plasticity
url https://doi.org/10.1038/s41598-024-83206-9
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