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

Nature Genetics volume 39, pages 251–258 (2007)Cite this article

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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Figure 1: Gradual removal of H3.1 and H3.2 in the XY body in spermatocytes during meiotic prophase I.
Figure 2: Timing of onset and duration of H3.1 and H3.2 removal.
Figure 3: Progressive incorporation of H3.3 at sex chromosomes and autosomes during mouse meiotic prophase.
Figure 4: Dynamics in histone lysine-methylation patterns in relation to nucleosomal exchange at the XY body (a–d).
Figure 5: Localization of H3.1 and H3.2 in asynapsed autosomal chromatin of the T(1;13)70H and T(1;13)Wa translocation–containing spermatocytes (a–g) and Sycp1−/− spermatocytes (h).

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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

  1. Maud Giele

    Present address: Orthopedic Research Laboratory, Department of Orthopedics, Radboud University Nijmegen Medical Centre, P.O. Box 9101, 6500 HB, Nijmegen, The Netherlands

  2. Pawel Pelczar

    Present address: Institute of Laboratory Animal Science, University of Zürich, Sternwartstrasse 6, CH 8091, Zürich, Switzerland

  3. Godfried W van der Heijden and Alwin A H A Derijck: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Obstetrics and Gynaecology, Radboud University Nijmegen Medical Centre, P.O. Box 9101, Nijmegen, 6500 HB, 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, Basel, CH 4058, 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, Nijmegen, 6525 GA, The Netherlands

    Derick G Wansink

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

    Johan van der Vlag

  5. Orthopedic Research Laboratory, Department of Orthopedics, Radboud University Nijmegen Medical Centre, P.O. Box 9101, 6500 HB, Nijmegen, The Netherlands

    Pawel Pelczar

  6. Institute of Laboratory Animal Science, University of Zürich, Sternwartstrasse 6, CH 8091, Zürich, Switzerland

    Maud Giele

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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.

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Supplementary information

Supplementary Fig. 1

Temporary reduction of nucleosome density in the XY body. (PDF 828 kb)

Supplementary Fig. 2

Generation and characterization of H3.3-V5 and H3.1-HA transgenic lines. (PDF 157 kb)

Supplementary Fig. 3

Patterns of H3K4, H3K9, H3K27 and H4K20 methylation during prophase I. (PDF 2158 kb)

Supplementary Fig. 4

Schematic overview of timing of events. (PDF 390 kb)

Supplementary Table 1

Presence of HirA in XY chromatin during prophase I. (PDF 23 kb)

Supplementary Table 2

Duration of H3.1/H3.2 loss and positioning in prophase I. (PDF 30 kb)

Supplementary Table 3

Frequencies of H3.3-V5 incorporation into sex chromosomes during subsequent stages of meiotic prophase and in round spermatids. (PDF 34 kb)

Supplementary Table 4

Histone H3.1/H3.2 in T70H/T1Wa and Sycp−/− spermatocytes. (PDF 25 kb)

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van der Heijden, G., Derijck, A., Pósfai, E. et al. Chromosome-wide nucleosome replacement and H3.3 incorporation during mammalian meiotic sex chromosome inactivation. Nat Genet 39, 251–258 (2007). https://doi.org/10.1038/ng1949

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