Letter | Published:

Maternal Rnf12/RLIM is required for imprinted X-chromosome inactivation in mice

Nature volume 467, pages 977981 (21 October 2010) | Download Citation

Abstract

Two forms of X-chromosome inactivation (XCI) ensure the selective silencing of female sex chromosomes during mouse embryogenesis. Imprinted XCI begins with the detection of Xist RNA expression on the paternal X chromosome (Xp) at about the four-cell stage of embryonic development. In the embryonic tissues of the inner cell mass, a random form of XCI occurs in blastocysts that inactivates either Xp or the maternal X chromosome (Xm)1,2. Both forms of XCI require the non-coding Xist RNA that coats the inactive X chromosome from which it is expressed. Xist has crucial functions in the silencing of X-linked genes, including Rnf12 (refs 3, 4) encoding the ubiquitin ligase RLIM (RING finger LIM-domain-interacting protein). Here we show, by targeting a conditional knockout of Rnf12 to oocytes where RLIM accumulates to high levels, that the maternal transmission of the mutant X chromosome (Δm) leads to lethality in female embryos as a result of defective imprinted XCI. We provide evidence that in Δm female embryos the initial formation of Xist clouds and Xp silencing are inhibited. In contrast, embryonic stem cells lacking RLIM are able to form Xist clouds and silence at least some X-linked genes during random XCI. These results assign crucial functions to the maternal deposit of Rnf12/RLIM for the initiation of imprinted XCI.

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Acknowledgements

We thank V. Boyartchuk, T. Fazzio, E. Heard, P. Kaufman, O. Rando, D. Riethmacher and J. Sharp for advice and/or reagents, D. Kim for help in ES cell analysis, and J. Zhu for statistics. I.B. is a member of the University of Massachusetts DERC (DK32520). This work was supported by National Institutes of Health grants R01CA131158 (National Cancer Institute) and 5 P30 DK32520 (National Institute of Diabetes and Digestive and Kidney Diseases) to I.B., and GM053234 to J.B.L.

Author information

Author notes

    • Michael Bossenz
    •  & Young Chung

    These authors contributed equally to this work.

    • Michael Bossenz

    Present address: Institute for Biochemistry and Cell Biology, University of Magdeburg, 39120 Magdeburg, Germany.

Affiliations

  1. Program in Gene Function and Expression, University of Massachusetts Medical School (UMMS), Worcester, Massachusetts 01605, USA

    • JongDae Shin
    • , Hong Ma
    • , Naoko Taniguchi-Ishigaki
    • , Xiaochun Zhu
    • , Baowei Jiao
    • , Michael R. Green
    •  & Ingolf Bach
  2. Program in Molecular Medicine, UMMS, Worcester, Massachusetts 01605, USA

    • Michael R. Green
    •  & Ingolf Bach
  3. Howard Hughes Medical Institute, UMMS, Worcester, Massachusetts 01605, USA

    • Xiaochun Zhu
    •  & Michael R. Green
  4. Department of Cell Biology, UMMS, Worcester, Massachusetts 01605, USA

    • Meg Byron
    • , Lisa L. Hall
    • , Stephen N. Jones
    •  & Jeanne B. Lawrence
  5. Centre for Molecular Neurobiology, University of Hamburg, 20246 Hamburg, Germany

    • Michael Bossenz
    •  & Irm Hermans-Borgmeyer
  6. Stem Cell and Regenerative Medicine International, Inc., Marlborough, Massachusetts 01752, USA

    • Young Chung
  7. CHA University, School of Medicine, Seoul 135-081, Korea

    • Young Chung

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Contributions

J.S. and I.B. conceived and designed the experiments. M.B., I.H.-B. and I.B. generated the floxed Rnf12 mice. J.S. and Y.C. established and analysed ES cell lines. J.S., M.B., H.M., M.B., N.T.-I., X.Z. and B.J. performed experiments. All authors analysed the data. I.B. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Ingolf Bach.

Supplementary information

PDF files

  1. 1.

    Supplementary Figures

    This file contains Supplementary Figures 1-14 with legends.

Videos

  1. 1.

    Supplementary Movie 1

    This movie shows a significant number of cells in the ICM - see Supplementary Information file for full legend.

  2. 2.

    Supplementary Movie 2

    This movie shows embryos develops Xist clouds - see Supplementary Information file for full legend.

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DOI

https://doi.org/10.1038/nature09457

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