Genomic imprinting recapitulated in the human beta-globin locus.
A subset of genes in mammals are subject to genomic imprinting. The mouse H19 gene, for example, is active only when maternally inherited and the neighboring Igf2 gene is paternally expressed. This imprinted expression pattern is regulated by the imprinting control region (ICR) upstream of the H19 gene. A maternally inherited H19 ICR inhibits Igf2 gene activation by the downstream enhancer due to its insulator function while it suppresses H19 gene transcription by promoter DNA methylation when paternally inherited. These parent-of-origin specific functions depend on the allele-specific methylation of the ICR DNA, which is established during gametogenesis. Therefore, the ICR may also function as a landmark for epigenetic modifications. To examine whether the ICR confers these activities autonomously, we introduced a 2.9-kbp ICR-containing DNA fragment into a human beta-globin yeast artificial chromosome at the 3' end of the locus control region and established transgenic mouse lines. Expression of all of the beta-like globin genes was higher when the transgene was paternally inherited. In accord with this result, transgenic ICR DNA from nucleated erythrocytes was more heavily methylated when paternally transmitted. Chromatin immunoprecipitation assays confirmed that CCCTC binding factor is preferentially recruited to the maternal transgenic ICR in vivo. Surprisingly however, the parent-of-origin specific methylation pattern was not observed in germ cell DNA in testis, demonstrating that methylation was established after fertilization. Thus, the ICR autonomously recapitulated imprinting within the normally nonimprinted transgenic beta-globin gene locus, but the temporal establishment of imprinting methylation differs from that at the endogenous Igf2/H19 locus.
Tanimoto
Keiji
K
Graduate School of Life and Environmental Sciences, Center for Tsukuba Advanced Research Alliance, University of Tsukuba, Tsukuba, Ibaraki 305-8577, Japan. keiji@tara.tsukuba.ac.jp
Shimotsuma
Motoshi
M
Matsuzaki
Hitomi
H
Omori
Akane
A
Bungert
Jörg
J
Engel
James Douglas
JD
Fukamizu
Akiyoshi
A
D000818:Animals; D047369:Chromatin Immunoprecipitation; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics; D003001:Cloning, Molecular; D019175:DNA Methylation; D004912:Erythrocytes / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation / Q000235:genetics*; D018392:Genomic Imprinting / Q000235:genetics*; D005914:Globins / Q000235:genetics*; D006801:Humans; D040582:Inheritance Patterns / Q000235:genetics; D007335:Insulin-Like Growth Factor II; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D011506:Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013737:Testis / Q000378:metabolism; D015533:Transcriptional Activation
C035170:IGF2 protein, human; D011506:Proteins; D007335:Insulin-Like Growth Factor II; D005914:Globins