Molecular coupling of Tsix regulation and pluripotency (original) (raw)

Nature volume 468, pages 457–460 (2010)Cite this article

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Abstract

The reprogramming of X-chromosome inactivation during the acquisition of pluripotency in vivo and in vitro1 is accompanied by the repression of Xist2, the trigger of X-inactivation3, and the upregulation of its antisense counterpart Tsix4. We have shown that key factors supporting pluripotency—Nanog, Oct4 and Sox2—bind within Xist intron 1 in undifferentiated embryonic stem cells (ESC) to repress Xist transcription5. However, the relationship between transcription factors of the pluripotency network and Tsix regulation has remained unclear5,6. Here we show that Tsix upregulation in embryonic stem cells depends on the recruitment of the pluripotent marker Rex1, and of the reprogramming-associated factors Klf4 and c-Myc, by the DXPas34 minisatellite associated with the Tsix promoter. Upon deletion of DXPas34, binding of the three factors is abrogated and the transcriptional machinery is no longer efficiently recruited to the Tsix promoter. Additional analyses including knockdown experiments further demonstrate that Rex1 is critically important for efficient transcription elongation of Tsix. Hence, distinct embryonic-stem-cell-specific complexes couple X-inactivation reprogramming and pluripotency, with Nanog, Oct4 and Sox2 repressing Xist to facilitate the reactivation of the inactive X, and Klf4, c-Myc and Rex1 activating Tsix to remodel Xist chromatin7,8,9,10 and ensure random X-inactivation upon differentiation1. The holistic pattern of Xist/Tsix regulation by pluripotent factors that we have identified suggests a general direct governance of complex epigenetic processes by the machinery dedicated to pluripotency.

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Acknowledgements

We thank P. Clerc for discussions. P.N. was initially supported by recurrent funding from the Institut Pasteur and then by the Royal Society (a Newton International Fellowship). P.A. was supported by recurrent funding from the Centre National de la Recherche Scientifique and the Institut Pasteur, contracts 05-JCJC-0166-01 and 07-BLAN-0047-01 from the Agence Nationale de la Recherche and funding from the EU Epigenome Network of Excellence. C.R. was supported by the INSERM ‘Avenir’ programme and by the European Research Council ‘starting grant’ programme. Research in I.C.’s laboratory was supported by the Wellcome Trust, by the EU Framework 7 project “EuroSyStem” and by a Medical Research Council studentship (to N.F.).

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Authors and Affiliations

  1. Unité de Génétique Moléculaire Murine, URA 2578, Institut Pasteur, 75724 Paris Cedex 15, France,
    Pablo Navarro, Andrew Oldfield, Julie Legoupi, Agnès Dubois, Mikael Attia, Claire Rougeulle & Philip Avner
  2. Medical Research Council (MRC) Centre Development in Stem Cell Biology, Institute for Stem Cell Research, School of Biological Sciences, University of Edinburgh, Edinburgh, EH9 3JQ, UK
    Pablo Navarro, Nicola Festuccia & Ian Chambers
  3. UMR 7216 Epigénétique et Destin Cellulaire, CNRS/Université Paris Diderot, Case 7042, 75205 Paris Cedex 13, France,
    Andrew Oldfield & Claire Rougeulle
  4. Departamento de Anatomía y Embriología, ARAID Foundation and Instituto Aragonés de Ciencias de la Salud, Facultad de Veterinaria, Zaragoza, 50013, Spain
    Jon Schoorlemmer

Authors

  1. Pablo Navarro
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  2. Andrew Oldfield
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  3. Julie Legoupi
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  4. Nicola Festuccia
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  5. Agnès Dubois
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  6. Mikael Attia
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  7. Jon Schoorlemmer
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  8. Claire Rougeulle
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  9. Ian Chambers
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  10. Philip Avner
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Contributions

P.N. conceived the study, designed, performed and analysed the experiments, and co-wrote the manuscript. A.O., J.L., N.F., M.A. and A.D. provided technical help. C.R., I.C. and P.A. provided financial and conceptual support. P.A. conceived the study and co-wrote the manuscript. All authors read and approved the final manuscript.

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Correspondence toPablo Navarro or Philip Avner.

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Navarro, P., Oldfield, A., Legoupi, J. et al. Molecular coupling of Tsix regulation and pluripotency.Nature 468, 457–460 (2010). https://doi.org/10.1038/nature09496

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Editorial Summary

Tsix regulation and pluripotency

Reprogramming of X-chromosome inactivation during the acquisition of pluripotency is accompanied by repression of Xist, the trigger of X-inactivation, and by upregulation of its antisense counterpart Tsix, which triggers resetting of the Xist locus. In undifferentiated embryonic stem cells, key transcription factors that support pluripotency — Nanog, Oct4 and Sox2 — repress Xist transcription. Here, upregulation of Tsix in embryonic stem cells is shown to depend on a different subset of pluripotency factors — Rex1, Klf4 and c-Myc. Therefore, two distinct embryonic-stem-cell-specific complexes couple reprogramming of X-inactivation to pluripotency.