Human Molecular Genetics Advance Access originally published online on August 4, 2004
Human Molecular Genetics 2004 13(19):2233-2245; doi:10.1093/hmg/ddh244
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Human Molecular Genetics, Vol. 13, No. 19 © Oxford University Press 2004; all rights reserved
Promoter-restricted histone code, not the differentially methylated DNA regions or antisense transcripts, marks the imprinting status of IGF2R in human and mouse
Medical Service, Veteran Affairs Palo Alto Health Care System and Department of Medicine, Stanford University, Palo Alto, CA 94304, USA
Received May 7, 2004; Revised July 8, 2004; Accepted July 20, 2004
Imprinting of the mouse Igf2r depends upon an intronic differentially methylated DNA region (DMR) and the presence of the Air antisense transcript. However, biallelic expression of mouse Igf2r in brain occurs despite the presence of Air, and biallelic expression of human IGF2R in peripheral tissues occurs despite the presence of an intronic DMR. We examined histone modifications throughout the mouse and human Igf2r/IGF2R using chromatin immuno-precipitation (ChIP) assays in combination with quantitative real time PCR. Methylation of Lys4 and Lys9 of histone H3 in the promoter regions marks the active and silenced alleles, respectively. We measured di- and tri-methyl Lys4 and Lys9 across the Igf2r and Air promoters. While both di- and tri-methyl Lys4 marked the active Igf2r and the active Air allele, tri-methyl Lys9, but not di-methyl Lys9, marked the suppressed Air allele. We show here that enrichment of parental allele-specific histone modifications in the promoter region, rather than the presence of DNA methylation or antisense transcription, correctly identifies the tissue- and species- specific imprinting status of Igf2r/IGF2R. We discuss these findings in light of recent progress in identifying specific components of the epigenetic marks in imprinted genes.
* To whom correspondence should be addressed at: Stanford Medical School, 3801 Miranda Avenue, Palo Alto, CA 94304, USA. Tel: +1 6504935000 ext. 63185; Fax: +1 6508568024; Email: thanhvu{at}stanford.edu
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