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Originally published In Press as doi:10.1074/jbc.M105775200 on January 30, 2002

J. Biol. Chem., Vol. 277, Issue 14, 11728-11734, April 5, 2002
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Inhibition of Histone Deacetylases Alters Allelic Chromatin Conformation at the Imprinted U2af1-rs1 Locus in Mouse Embryonic Stem Cells*

Richard I. GregoryDagger §, Laura P. O'Neill, Tamzin E. Randall, Cecile Fournier||, Sanjeev KhoslaDagger **, Bryan M. Turner, and Robert FeilDagger ||Dagger Dagger

From the || Institute of Molecular Genetics, CNRS UMR-5535, IFR-24, 34293 Montpellier cedex 5, France, Dagger  The Babraham Institute, Cambridge CB2 4AT, United Kingdom, and the  Chromatin and Gene Expression Group, University of Birmingham Medical School, Birmingham B15 2TT, United Kingdom

Most loci that are regulated by genomic imprinting have differentially methylated regions (DMRs). Previously, we showed that the DMRs of the mouse Snrpn and U2af1-rs1 genes have paternal allele-specific patterns of acetylation on histones H3 and H4. To investigate the maintenance of acetylation at these DMRs, we performed chromatin immunoprecipitation on trichostatin-A (TSA)-treated and control cells. In embryonic stem (ES) cells and fibroblasts, brief (6-h) TSA treatment induces global hyperacetylation of H3 and H4. In ES cells only, TSA led to a selective increase in maternal acetylation at U2af1-rs1, at lysine 5 of H4 and at lysine 14 of H3. TSA treatment of ES cells did not affect DNA methylation or expression of U2af1-rs1, but was sufficient to increase DNase I sensitivity along the maternal allele to a level comparable with that of the paternal allele. In fibroblasts, TSA did not alter U2af1-rs1 acetylation, and the parental alleles retained their differential DNase I sensitivity. At Snrpn, no changes in acetylation were observed in the TSA-treated cells. Our data suggest that the mechanisms regulating histone acetylation at DMRs are locus and developmental stage-specific and are distinct from those effecting global levels of acetylation. Furthermore, it seems that the allelic U2af1-rs1 acetylation determines DNase I sensitivity/chromatin conformation.

* This work was supported by the Biotechnology and Biological Sciences Research Council (Studentship to R. I. G.), the Center National de la Recherche Scientifique (to R. F.), the Human Frontier Science Program (to R. F.), the Fondation pour la Recherche Médicale (to R. F.), and the Royal Society (Fellowship 516002 (to L. P. O.)).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AF309654.

§ Present address: Fox Chase Cancer Center, 7701 Burholme Ave., Philadelphia, PA 19111.

** Present address: Wellcome/CRC Inst. of Developmental Biology and Cancer Research, University of Cambridge, Cambridge CB2 1QR, United Kingdom.

Dagger Dagger To whom correspondence should be addressed. Tel.: 33-4-67613663; Fax: 33-4-67040231; E-mail: feil@igm.cnrs-mop.fr.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.


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