Phosphorylation of histone H3 serine 10 in early mouse embryos: Active phosphorylation at late S phase and differential effects of ZM447439 on first two embryonic

Department of Embryology, Institute of Zoology, Faculty of Biology, University of Warsaw, Warsaw, Poland.
Cell cycle (Georgetown, Tex.) (Impact Factor: 4.57). 12/2010; 9(23):4674-87. DOI: 10.4161/cc.9.23.14023
Source: PubMed


Cell division in mammalian cells is regulated by Aurora kinases. The activity of Aurora A is indispensable for correct function of centrosomes and proper spindle formation, while Aurora B for chromosome biorientation and separation. Aurora B is also responsible for the phosphorylation of histone H3 serine 10 (H3S10Ph) from G2 to metaphase. Data concerning the Aurora B activity and H3S10Ph in embryonic cells are limited to primordial and maturing oocytes and advanced pronuclei in zygotes. In the present study we have analyzed H3S10Ph in 1- and 2-cell mouse embryos. We show that H3S10 remains phosphorylated at anaphase and telophase of the second meiotic division, as well as during the anaphase and telophase of the first and second embryonic mitoses. At late G1 H3S10 is dephosphorylated and subsequently phosphorylated de novo at late S phase of the first and second cell cycle. These results show that the H3S10 phosphorylation/dephosphorylation cycle in embryonic cells is different than in somatic cells. The behaviour of thymocyte G0 nuclei introduced into ovulated oocytes and early 1-cell parthenogenotes confirms that kinases responsible for de novo H3S10 phosphorylation, most probably Aurora B, are active until G1 of the first cell cycle of mouse embryo. The inhibition of Aurora kinases by ZM447439 caused abnormalities both in the first and second mitoses. However, the disturbances in each division differed, suggesting important differences in the control of these mitoses. In ZM447439-treated mitotic zygotes Mad2 protein remained continuously present on kinetochores, what confirmed that spindle checkpoint remained active.

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    • "During interphase , binding of HP1 to di-or trimethylated H3K9 maintains a constitutive heterochromatin status. When cells enter mitosis, however, phosphorylation of H3S10, acetylation of H3K14, and possibly methylation of H3K4, hinder methylation of H3K9, causing HP1 to be released from the histone (Fischle et al., 2005; Hirota et al., 2005; Teperek-Tkacz et al., 2010), bringing a change to the higher structure of chromatin. This mechanism , which appears to be fundamental to the role of H3S10 phosphorylation in chromosome condensation during mitosis, has been suggested to apply as well to H3S10 phosphorylation in the promoters of certain genes to regulate transcription during interphase (Winter et al., 2008). "
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    ABSTRACT: Aryl hydrocarbon receptor (AHR) activation by xenobiotic ligands such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is key to their toxicity. Following activation and nuclear translocation, AHR heterodimerizes with the AHR nuclear translocator (ARNT) and binds to AHR response elements (AhREs) in the enhancer of target genes, of which Cyp1a1 is the prototype. Previously, we showed that concomitant with AHR binding, histone H3 in the Cyp1a1 enhancer-promoter AhRE cluster became phosphorylated in serine-10 (H3S10), suggesting that the ligand-activated AHR recruited one or more kinases to the enhancer chromatin to phosphorylate this residue. To test this hypothesis, we used mouse hepatoma Hepa-1c1c7 cells and their c35 mutant derivative, lacking a functional AHR, to search for candidate kinases that would phosphorylate H3S10 in an AHR dependent manner. Using chromatin immunoprecipitation with antibodies to a comprehensive set of protein kinases, we identified three kinases, IKKα, MSK1 and MSK2, whose binding to the Cyp1a1 enhancer was significantly increased by TCDD in Hepa-1c1c7 cells and absent in control c35 cells. Complexes of AHR, ARNT and IKKα could be co-immunoprecipitated from nuclei of TCDD treated Hepa-1c1c7 cells, and shRNA-mediated IKKα knockdown inhibited both H3S10 phosphorylation in the Cyp1a1 enhancer and the induction of Cyp1a1, Aldh3a1 and Nqo1 in TCDD-treated cells. We conclude that AHR recruits IKKα to the promoter of its target genes and that AHR-mediated H3S10 phosphorylation is a key epigenetic requirement for induction of AHR targets. Given the role of H3S10ph in regulation of chromosome condensation, AHR-IKKα crosstalk may be a mediator of chromatin remodeling by environmental agents.
    Toxicological Sciences 02/2014; 139(1). DOI:10.1093/toxsci/kfu027 · 3.85 Impact Factor
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    • "For this purpose, embryos were obtained from superovulated F1(C57BL/6xCBA/Tar) females mated with F1 males and the timing of each cell cycle progression was estimated based on the time that passed from hCG injection (in hours post hCG). Such precise timing for this particular strain of mice has been established previously (Teperek-Tkacz et al., 2010) and confirmed (Meglicki et al., 2012). To exclude possible artifacts resulting from using a different mouse strain or a different method of obtaining embryos, we first determined whether localization of LIN28 in embryos derived from superovulated F1 and spontaneously ovulated CD1 mice was similar. "
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    ABSTRACT: The maternal nucleolus is required for proper activation of the embryonic genome (EGA) and early embryonic development. Nucleologenesis is characterized by the transformation of a nucleolar precursor body (NPB) to a mature nucleolus during preimplantation development. However, the function of NPBs and the involved molecular factors are unknown. We uncover a novel role for the pluripotency factor LIN28, the biological significance of which was previously demonstrated in the reprogramming of human somatic cells to induced pluripotent stem (iPS) cells. Here, we show that LIN28 accumulates at the NPB and the mature nucleolus in mouse preimplantation embryos and embryonic stem cells (ESCs), where it colocalizes with the nucleolar marker B23 (nucleophosmin 1). LIN28 has nucleolar localization in non-human primate (NHP) preimplantation embryos, but is cytoplasmic in NHP ESCs. Lin28 transcripts show a striking decline before mouse EGA, whereas LIN28 protein localizes to NPBs at the time of EGA. Following knockdown with a Lin28 morpholino, the majority of embryos arrest between the 2- and 4-cell stages and never develop to morula or blastocyst. Lin28 morpholino-injected embryos arrested at the 2-cell stage were not enriched with nucleophosmin at presumptive NPB sites, indicating that functional NPBs were not assembled. Based on these results, we propose that LIN28 is an essential factor of nucleologenesis during early embryonic development.
    Development 12/2012; 139(24):4514-23. DOI:10.1242/dev.083279 · 6.46 Impact Factor
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    • "Some research groups have investigated this epigenetic modification during early mouse embryogenesis. Their reports suggest that H3S10P is linked to pericentromeric heterochromatin (Huang et al., 2007; Teperek-Tkacz et al., 2010; Wang et al., 2006). Recently, we have shown by immunofluorescence in situ hybridization (immuno-FISH) using specific pericentromeric heterochromatin probes that H3S10P indeed marks constitutive heterochromatin during interphase until the four-cell stage in mouse embryos (Ribeiro-Mason et al., submitted). "
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    ABSTRACT: Somatic cell nuclear transfer (SCNT) is the injection of a donor nucleus into an enucleated egg. Despite the use of this technology for many years in research, it is still quite inefficient. One of the causes for this is thought to be incorrect or incomplete genome reprogramming. Embryos produced by nuclear transfer (cloned embryos) very often present abnormal epigenetic signatures and irregular chromatin reorganization. Of these two issues, the issue of chromatin rearrangements within the nuclei after transfer is the least studied. It is known that cloned embryos often present pericentromeric heterochromatin clumps very similar to the chromocenters structures present in the donor nuclei. Therefore, it is believed that the somatic nuclear configuration of donor nuclei, especially that of the chromocenters, is not completely lost after nuclear transfer, in other words, not well reprogrammed. To further investigate pericentromeric heterochromatin reorganization after nuclear transfer, we decided to study its rearrangements in cumulus-derived clones using several related epigenetic markers such as H3S10P, H3K9me3, and the double marker H3K9me3S10P. We observed that two of these markers, H3S10P and H3K9me3S10P, are the ones found on the part of the pericentromeric heterochromatin that is remodeled correctly, resembling exactly the embryonic heterochromatin configuration of naturally fertilized embryos. Conversely, H3K9me3 and heterochromatin protein 1 beta (HP1β)-associated protein were also detected in the perinuclear clumps of heterochromatin, making obvious the maintenance of the somatic epigenetic signature within these nuclear regions. Our results demonstrate that H3S10P and H3K9me3S10P could be good candidates for evaluating heterochromatin reorganization following nuclear reprogramming.
    07/2012; 14(4):283-94. DOI:10.1089/cell.2011.0071
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