Genome wide identification of promoter binding sites for H4K12ac in human sperm and its relevance for early embryonic development

• Published in: Epigenetics 2012-09, Vol.7 (9), p.1057-1070
• Main Authors: Paradowska, Agnieszka S., Miller, David, Spiess, Andrej-Nikolai, Vieweg, Markus, Cerna, Martina, Dvorakova-Hortova, Katerina, Bartkuhn, Marek, Schuppe, Hans-Christian, Weidner, Wolfgang, Steger, Klaus
• Format: Article
• Language: English
• Subjects: Acetylation; Animals; Binding; Binding Sites; Biology; Bioscience; Blastocyst - metabolism; Calcium; Cancer; Cell; Chromatin - genetics; Chromatin - metabolism; chromatin immunoprecipitation; Cycle; Embryonic Development - genetics; Epigenesis, Genetic; Female; Gene Expression Regulation, Developmental; Genes, Developmental; Genome, Human; H4K12ac; HG18 promoter microarray; Histones - metabolism; Humans; Landes; Male; Membrane Proteins - genetics; Membrane Proteins - metabolism; Mice; Mice, Inbred C57BL; mouse pronucleus; Organogenesis; parthenotes; Promoter Regions, Genetic; Protein Processing, Post-Translational; Proteins; Research Paper; RNA, Messenger - biosynthesis; sperm histone code; sperm mRNA; Spermatozoa - metabolism; Transcription Factors - genetics; Transcription Factors - metabolism
• ISSN: 1559-2294; 1559-2308
• DOI: 10.4161/epi.21556

Sperm chromatin reveals two characteristic features in that protamines are the predominant nuclear proteins and remaining histones are highly acetylated. Histone H4 acetylated at lysine 12 (H4K12ac) is localized in the post-acrosomal region, while protamine-1 is present within the whole nucleus. Chromatin immunoprecipitation in combination with promoter array analysis allowed genome-wide identification of H4K12ac binding sites. Previously, we reported enrichment of H4K12ac at CTCF binding sites and promoters of genes involved in developmental processes. Here, we demonstrate that H4K12ac is enriched predominantly between ± 2 kb from the transcription start site. In addition, we identified developmentally relevant H4K12ac-associated promoters with high expression levels of their transcripts stored in mature sperm. The highest expressed mRNA codes for testis-specific PHD finger protein-7 (PHF7), suggesting an activating role of H4K12ac in the regulatory elements of this gene. H4K12ac-associated genes revealed a weak correlation with genes expressed at 4-cell stage human embryos, while 23 H4K12ac-associated genes were activated in 8-cell embryo and 39 in the blastocyst. Genes activated in 4-cell embryos are involved in gene expression, histone fold and DNA-dependent transcription, while genes expressed in the blastocyst were classified as involved in developmental processes. Immunofluorescence staining detected H4K12ac from the murine male pronucleus to early stages of embryogenesis. Aberrant histone acetylation within developmentally important gene promoters in infertile men may reflect insufficient sperm chromatin compaction, which may result in inappropriate transfer of epigenetic information to the oocyte.