Systematic genetic and proteomic screens during gametogenesis identify H2BK34 methylation as an evolutionary conserved m
- PDF / 2,062,918 Bytes
- 23 Pages / 595.276 x 790.866 pts Page_size
- 69 Downloads / 175 Views
Epigenetics & Chromatin Open Access
RESEARCH
Systematic genetic and proteomic screens during gametogenesis identify H2BK34 methylation as an evolutionary conserved meiotic mark Marion Crespo1,2†, Lacey J. Luense3,4†, Marie Arlotto1,2,5†, Jialei Hu3,4, Jean Dorsey3,4, Encar García‑Oliver1,6, Parisha P. Shah3,4, Delphine Pflieger1,2, Shelley L. Berger3,4 and Jérôme Govin1,5*
Abstract Background: Gametes are highly differentiated cells specialized to carry and protect the parental genetic informa‑ tion. During male germ cell maturation, histone proteins undergo distinct changes that result in a highly compacted chromatin organization. Technical difficulties exclude comprehensive analysis of precise histone mutations during mammalian spermatogenesis. The model organism Saccharomyces cerevisiae possesses a differentiation pathway termed sporulation which exhibits striking similarities to mammalian spermatogenesis. This study took advantage of this yeast pathway to first perform systematic mutational and proteomics screens on histones, revealing amino acid residues which are essential for the formation of spores. Methods: A systematic mutational screen has been performed on the histones H2A and H2B, generating ~ 250 mutants using two genetic backgrounds and assessing their ability to form spores. In addition, histones were purified at key stages of sporulation and post-translational modifications analyzed by mass spectrometry. Results: The mutation of 75 H2A H2B residues affected sporulation, many of which were localized to the nucleosome lateral surface. The use of different genetic backgrounds confirmed the importance of many of the residues, as 48% of yeast histone mutants exhibited impaired formation of spores in both genetic backgrounds. Extensive proteomic analysis identified 67 unique post-translational modifications during sporulation, 27 of which were previously unre‑ ported in yeast. Furthermore, 33 modifications are located on residues that were found to be essential for efficient sporulation in our genetic mutation screens. The quantitative analysis of these modifications revealed a massive deacetylation of all core histones during the pre-meiotic phase and a close interplay between H4 acetylation and methylation during yeast sporulation. Methylation of H2BK37 was also identified as a new histone marker of meiosis and the mouse paralog, H2BK34, was also enriched for methylation during meiosis in the testes, establishing conser‑ vation during mammalian spermatogenesis. Conclusion: 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.
*Correspondence: [email protected] † Marion Crespo, Lacey J. Luense and Marie Arlotto contributed equally to this work 1 Univ. Grenoble Alpes, CEA, Inserm, IRIG-BGE, 38000 Grenoble, France Full list of author information is available at the end of the article
Introduction Gametes are highly specialized cells dedicated to the r
Data Loading...
