Abstract
Histone phosphorylation is sometimes associated with mitosis and meiosis. We have recently identified a phosphorylation of the 127th threonine on TH2A (pTH2A), a germ cell-specific H2A variant, in condensed spermatids and mitotic early preimplantation embryos of mice. Here, we further report the existence of pTH2A at the centromeres in metaphase I spermatocytes and oocytes. Moreover, we identified Haspin, a known kinase for the 3rd threonine on H3, is responsible for pTH2A in vivo. In contrast to the severe meiotic defect in oocytes treated with a Haspin inhibitor, pTH2A-deficient mice, in which the 127th threonine was replaced by alanine, maintained the fertility and exhibited no obvious defect in both oocytes and spermatogenesis. Interestingly, pTH2A was significantly decreased in aged oocytes, suggesting that its accumulation is regulated by centromeric cohesins. Collectively, our study proposes a new set of kinase-histone pair at meiotic centromere, which is highly coordinated during meiosis.
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Acknowledgments
We would like to give a special thanks to the members of MBL antibody production team for generating anti-pTH2A antibody. We also thank Dr. Y. Makino and Mr. I. Tsuchiya from the laboratory of Pathology, Drs. H. Shibuya, Y. Tanno, and T. Sakuno from the laboratory of Chromosome Dynamics at the Institute of Molecular and Cellular Biosciences, the University of Tokyo, Drs. Tokoro and Yamagata from Kinki University, and Dr. J Lee from Kobe University for helpful advice and technical assistance, Drs. T. Shinagawa and S. Ishii from the RIKEN Institute for kindly providing the anti-TH2A antibody, and Drs. H. Tanaka and H. Kurumizaka from Waseda University for kindly providing the recombinant TH2A.
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M.H. and Y.O. designed the study, and M.H., J. K., E. I., Y. F., and Y. O. performed experiments. M.H. and Y.O. wrote the paper with helpful discussion from H. T. and Y.W.
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All experimental procedures involving mice were approved by the Animal Experiment Ethics Committees at the Institute of Molecular and Cellular Biosciences, The University of Tokyo (Experiment# 2611, 2710, 2808), and the experiments were performed precisely in accordance with the manual provided by the Life Science Research Ethics and Safety, the University of Tokyo.
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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed; all experimental procedures involving mice were approved by the Animal Experiment Ethics Committees at the Institute of Molecular and Cellular Biosciences, The University of Tokyo (Experiment# 2611, 2710, 2808), and the experiments were performed precisely in accordance with the manual provided by the Life Science Research Ethics and Safety, the University of Tokyo.
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This article does not contain any studies with human participants performed by any of the authors.
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This work was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) in Japan (22125007 and 15H05976 to Y.O.). M.H. is a Research Fellow (DC2) for Young Scientists, JSPS (14J10178). This work was also partially supported by the Platform Project for Supporting in Drug Discovery and Life Science Research (Platform for Drug Discovery, Informatics, and Structural Life Science) from Japan Agency for Medical Research and Development (AMED).
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Hada, M., Kim, J., Inoue, E. et al. TH2A is phosphorylated at meiotic centromere by Haspin. Chromosoma 126, 769–780 (2017). https://doi.org/10.1007/s00412-017-0638-5
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DOI: https://doi.org/10.1007/s00412-017-0638-5