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Chromosome-wide nucleosome replacement and H3.3 incorporation during mammalian meiotic sex chromosome inactivation

Nature Genetics volume 39, pages 251258 (2007) | Download Citation

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Abstract

In mammalian males, the first meiotic prophase is characterized by formation of a separate chromatin domain called the sex body1. In this domain, the X and Y chromosomes are partially synapsed and transcriptionally silenced, a process termed meiotic sex-chromosome inactivation (MSCI)2,3. Likewise, unsynapsed autosomal chromatin present during pachytene is also silenced (meiotic silencing of unsynapsed chromatin, MSUC)2,4,5. Although it is known that MSCI and MSUC are both dependent on histone H2A.X phosphorylation mediated by the kinase ATR, and cause repressive H3 Lys9 dimethylation4, the mechanisms underlying silencing are largely unidentified. Here, we demonstrate an extensive replacement of nucleosomes within unsynapsed chromatin, depending on and initiated shortly after induction of MSCI and MSUC. Nucleosomal eviction results in the exclusive incorporation of the H3.3 variant, which to date has primarily been associated with transcriptional activity. Nucleosomal exchange causes loss and subsequent selective reacquisition of specific histone modifications. This process therefore provides a means for epigenetic reprogramming of sex chromatin presumably required for gene silencing in the male mammalian germ line.

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Acknowledgements

We thank T. Jenuwein, A. Schulmeister, F. van Leeuwen, C. Heyting, P.B. Moens, P.D. Adams and H.G. Stunnenberg for providing antibody reagents; J.-F. Spetz, A. Kelly and M. Puschendorf for their help in the generation and initial characterization of H3.1-HA and H3.3-V5 transgenic mice; C. Heyting, A. Pastink and E. de Boer for male meiotic preparations of Sycp1−/− knockout mice; and W.M. Baarends, C. Logie and P.J. Wang for critical reading of the manuscript. Research in the laboratory of A.H.F.M.P. is supported by the Novartis Research Foundation and the NoE network “The Epigenome” (LSHG-CT-2004-503433).

Author information

Author notes

    • Maud Giele
    •  & Pawel Pelczar

    Current addresses: Orthopedic Research Laboratory, Department of Orthopedics, Radboud University Nijmegen Medical Centre, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands (M.G.) and Institute of Laboratory Animal Science, University of Zürich, Sternwartstrasse 6, CH 8091 Zürich, Switzerland (P.P.).

    • Godfried W van der Heijden
    •  & Alwin A H A Derijck

    These authors contributed equally to this work.

Affiliations

  1. Department of Obstetrics and Gynaecology, Radboud University Nijmegen Medical Centre, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands.

    • Godfried W van der Heijden
    • , Alwin A H A Derijck
    • , Maud Giele
    • , Liliana Ramos
    •  & Peter de Boer
  2. Friedrich Miescher Institute for Biomedical Research, Maulbeerstrasse 66, CH 4058 Basel, Switzerland.

    • Eszter Pósfai
    • , Pawel Pelczar
    •  & Antoine H F M Peters
  3. Department of Cell Biology, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, Geert Grooteplein 26, 6525 GA Nijmegen, The Netherlands.

    • Derick G Wansink
  4. Nephrology Research Laboratory, Division of Nephrology, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, Geert Grooteplein 26, 6525 GA Nijmegen, The Netherlands.

    • Johan van der Vlag

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Contributions

G.W.v.d.H., A.A.H.A.D., E.P., A.H.F.M.P. and P.d.B. conceived and designed the experiments. G.W.v.d.H., A.A.H.A.D., E.P. and M.G. performed the experiments. G.W.v.d.H., A.A.H.A.D., E.P., A.H.F.M.P. and P.d.B. analyzed the data. P.P. generated histone-tagged transgenic mice. L.R. was responsible for human material. J.v.d.V. contributed H3.1- and H3.2-specific antibodies. G.W.v.d.H., A.A.H.A.D., D.G.W., J.v.d.V., A.H.F.M.P. and P.d.B. contributed to the writing of the manuscript. J.V.d.V. and A.H.F.M.P. contributed equally to this work.

Competing interests

The authors declare no competing financial interests.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    Temporary reduction of nucleosome density in the XY body.

  2. 2.

    Supplementary Fig. 2

    Generation and characterization of H3.3-V5 and H3.1-HA transgenic lines.

  3. 3.

    Supplementary Fig. 3

    Patterns of H3K4, H3K9, H3K27 and H4K20 methylation during prophase I.

  4. 4.

    Supplementary Fig. 4

    Schematic overview of timing of events.

  5. 5.

    Supplementary Table 1

    Presence of HirA in XY chromatin during prophase I.

  6. 6.

    Supplementary Table 2

    Duration of H3.1/H3.2 loss and positioning in prophase I.

  7. 7.

    Supplementary Table 3

    Frequencies of H3.3-V5 incorporation into sex chromosomes during subsequent stages of meiotic prophase and in round spermatids.

  8. 8.

    Supplementary Table 4

    Histone H3.1/H3.2 in T70H/T1Wa and Sycp−/− spermatocytes.

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DOI

https://doi.org/10.1038/ng1949

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