期刊
NUCLEIC ACIDS RESEARCH
卷 38, 期 15, 页码 4929-4945出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkq200
关键词
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资金
- Ministry of Education, Culture, Sports, Science and Technology of Japan [19390423, 20017003, 21028003]
- Japan Science and Technology Agency (JST)
- Takeda Science Foundation
- Children's Cancer Association of Japan
- National Institute of Genetics (NIG)
- Mitsubishi Foundation
- Smoking Research Foundation
- Nestle Nutrition Council Japan
- Grants-in-Aid for Scientific Research [21028003, 20062010, 20017003, 19390423] Funding Source: KAKEN
The parent-of-origin specific expression of imprinted genes relies on DNA methylation of CpG-dinucleotides at differentially methylated regions (DMRs) during gametogenesis. To date, four paternally methylated DMRs have been identified in screens based on conventional approaches. These DMRs are linked to the imprinted genes H19, Gtl2 (IG-DMR), Rasgrf1 and, most recently, Zdbf2 which encodes zinc finger, DBF-type containing 2. In this study, we applied a novel methylated-DNA immunoprecipitation-on-chip (meDIP-on-chip) method to genomic DNA from mouse parthenogenetic- and androgenetic-derived stem cells and sperm and identified 458 putative DMRs. This included the majority of known DMRs. We further characterized the paternally methylated Zdbf2/ZDBF2 DMR. In mice, this extensive germ line DMR spanned 16 kb and possessed an unusual tripartite structure. Methylation was dependent on DNA methyltransferase 3a (Dnmt3a), similar to H19 DMR and IG-DMR. In both humans and mice, the adjacent gene, Gpr1/GPR1, which encodes a G-protein-coupled receptor 1 protein with transmembrane domain, was also imprinted and paternally expressed. The Gpr1-Zdbf2 domain was most similar to the Rasgrf1 domain as both DNA methylation and the actively expressed allele were in cis on the paternal chromosome. This work demonstrates the effectiveness of meDIP-on-chip as a technique for identifying DMRs.
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