Molecular coupling of Tsix regulation and pluripotency

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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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Figure 1: DXPas34 orchestrates Rex1, Klf4 and c-Myc recruitment to the Tsix 5′ region in pluripotent ESC.
Figure 2: Developmentally induced loss of Rex1, Klf4 and c-Myc binding correlates with Tsix repression.
Figure 3: Rex1 is required for efficient elongation of Tsix transcription.

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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.).

Author information

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.

Correspondence to Pablo 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) doi:10.1038/nature09496

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