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DNA Research Advance Access originally published online on January 11, 2006
DNA Research 2005 12(5):373-378; doi:10.1093/dnares/dsi013
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© The Author 2006. Kazusa DNA Research Institute
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org

XIST Repression in the Absence of DNMT1 and DNMT3B

Luciana R. Vasques1, Raquel Stabellini1, Fei Xue2, X. Cindy Tian2, Marina Soukoyan1 and Lygia V. Pereira1,*

1Depto Biologia and Centro de Estudos do Genoma Humano, Instituto de Biociências, Universidade de São Paulo São Paulo, SP 05508-900, Brazil
2Center for Regenerative Biology/Department of Animal Science, University of Connecticut Storrs, CT 06269, USA

X chromosome inactivation (XCI) in human and mice involves XIST/Xist gene expression from the inactive X (Xi) and repression from the active X (Xa). Repression of the XIST/Xist gene on the Xa has been associated with methylation of its 5' region. In mice, Dnmt1 has been shown to be involved in the methylation and transcriptional repression of Xist on Xa. We examined maintenance of XIST gene repression on Xa in HCT116 cell lines knockout for either DNMT1 or DNMT3B and for DNMT1 and DNMT3B simultaneously. Methylation of the XIST promoter and XIST transcriptional repression is sustained in DNMT1-, DNMT3B- and DNMT1/DNMT3B knockout cells. Despite global DNA demethylation, the double knockout cells present only partial demethylation of the XIST promoter, which is not sufficient for gene reactivation. In contrast, global DNA demethylation with 5-aza-2'-deoxycytidine leads to XIST expression. Therefore, in these human cells maintenance of XIST methylation is controlled differently than global genomic methylation and in the absence of both DNMT1 and DNMT3B.

Key words: X-chromosome inactivation; DNA-methyltransferase; XIST; epigenetic inheritance

*To whom correspondence should be addressed. Tel. +55-11-3091-7476, Fax. +55-11-3091-7553, E-mail: LPEREIRA{at}USP.BR

Communicated by Mitsuo Oshimura


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