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Vol. 12, Issue 12, 1860-1870, December 2002

LETTER
Domain Regulation of Imprinting Cluster in Kip2/Lit1 Subdomain on Mouse Chromosome 7F4/F5: Large-Scale DNA Methylation Analysis Reveals That DMR-Lit1 Is a Putative Imprinting Control Region

Hitomi Yatsuki,1 Keiichiro Joh,1 Ken Higashimoto,1 Hidenobu Soejima,1 Yuji Arai,2 Youdong Wang,1 Izuho Hatada,3 Yayoi Obata,3 Hiroko Morisaki,2 Zhongming Zhang,1 Tetsuji Nakagawachi,1 Yuji Satoh,1 and Tsunehiro Mukai1,4

1 Department of Biochemistry, Saga Medical School, Saga, Saga 849-8501, Japan; 2 Department of Bioscience, National Cardiovascular Center Research Institute, Fujishiro-dai, Suita, Osaka 565-8565, Japan; 3 Gene Research Center, Gunma University, Showa-machi, Maebashi 371-8511, Japan

Mouse chromosome 7F4/F5, where the imprinting domain is located, is syntenic to human 11p15.5, the locus for Beckwith-Wiedemann syndrome. The domain is thought to consist of the two subdomains Kip2 (p57kip2)/Lit1 and Igf2/H19. Because DNA methylation is believed to be a key factor in genomic imprinting, we performed large-scale DNA methylation analysis to identify the cis-element crucial for the regulation of the Kip2/Lit1 subdomain. Ten CpG islands (CGIs) were found, and these were located at the promoter sites, upstream of genes, and within intergenic regions. Bisulphite sequencing revealed that CGIs 4, 5, 8, and 10 were differentially methylated regions (DMRs). CGIs 4, 5, and 10 were methylated paternally in somatic tissues but not in germ cells. CGI8 was methylated in oocyte and maternally in somatic tissues during development. Parental-specific DNase I hypersensitive sites (HSSs) were found near CGI8. These data indicate that CGI8, called DMR-Lit1, is not only the region for gametic methylation but might also be the imprinting control region (ICR) of the subdomain.

4 Corresponding author.

12:1860-1870 ©2002 by Cold Spring Harbor Laboratory Press  ISSN 1088-9051/02 $5.00
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