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Reactivation of Х chromosome upon reprogramming leads to changes in the replication pattern and 5hmC accumulation

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Abstract

Once set, the inactive status of the X chromosome in female somatic cells is preserved throughout subsequent cell divisions. The inactive status of the X chromosome is characterized by many features, including late replication. In contrast to induced pluripotent stem cells (iPSCs) in mice, the X chromosome in human female iPSCs usually remains inactive after reprogramming of somatic cells to the pluripotent state, although recent studies point to the possibility of reactivation of the X chromosome. Here, we demonstrated that, during reprogramming, the inactive X chromosome switches from late to synchronous replication, with restoration of the transcription of previously silenced genes. This process is accompanied by accumulation of a new epigenetic mark or intermediate of the DNA demethylation pathway, 5-hydroxymethylcytosine (5hmC), on the activated X chromosome. Our results indicate that the active status of the X chromosome is better confirmed by early replication and the reappearance of 5hmC, rather than by appearance of histone marks of active chromatin, removal of histone marks of inactive chromatin, or an absence of XIST coating.

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Abbreviations

5hmC:

5-hydroxymethylcytosine

5mC:

5-methylcytosine

EdU:

5-ethynyl-2′-deoxyuridine

ESCs:

Embryonic stem cells

iPSCs:

Induced pluripotent stem cells

FISH:

Fluorescence in situ hybridization

Xa:

Active X chromosome

Xi:

Inactive X chromosome

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Acknowledgments

This research was supported by RFBR (11-04-01212a) and Ministry of Science and Education (GK 14.512.11.0025).

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Authors and Affiliations

  1. Vavilov Institute of General Genetics, Russian Academy of Sciences, 3, Gubkina St, Moscow, 119991, Russia

    Alexandra N. Bogomazova, Maria A. Lagarkova, Alexandra V. Panova, Evgueny D. Nekrasov & Sergey L. Kiselev

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  1. Alexandra N. Bogomazova
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  2. Maria A. Lagarkova
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  3. Alexandra V. Panova
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  4. Evgueny D. Nekrasov
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  5. Sergey L. Kiselev
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Corresponding author

Correspondence to Maria A. Lagarkova.

Additional information

Alexandra N. Bogomazova and Maria A. Lagarkova contributed equally to the publication.

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Bogomazova, A.N., Lagarkova, M.A., Panova, A.V. et al. Reactivation of Х chromosome upon reprogramming leads to changes in the replication pattern and 5hmC accumulation. Chromosoma 123, 117–128 (2014). https://doi.org/10.1007/s00412-013-0433-x

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  • DOI: https://doi.org/10.1007/s00412-013-0433-x

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