Shemer R, Kafri T, O'Connell A, Eisenberg S, Breslow J L, Razin A
Department of Cellular Biochemistry, Hebrew University Medical School, Jerusalem, Israel.
Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11300-4. doi: 10.1073/pnas.88.24.11300.
We report here a detailed study of developmental changes in the methylation status of specific sites in a single-copy tissue-specific gene, from the germ cell through the early embryo to adult tissues. Two sites at the 5' end of the mouse apolipoprotein AI gene were unmethylated in the ovulated unfertilized oocytes and methylated in the sperm. In contrast, a third site, located upstream of the gene, was methylated and a CpG island within the gene was unmethylated in both oocyte and sperm. The methylated sites, regardless of maternal or paternal origin, underwent demethylation in the early embryo (8-16 cells) and stayed unmethylated through the late blastocyst stage. During gastrulation, non-CpG island sites underwent methylation, followed by gradual demethylation at specific sites in tissues parallel to expression of the gene (liver and intestine). The formation of the mature tissue-specific methylation pattern of the apolipoprotein AI gene, therefore, involves the following three major events: (i) erasure of the germ-cell methylation pattern (at the 8- to 16-cell stage), (ii) formation of a new methylation pattern by de novo methylation of non-CpG island sites (during gastrulation), and (iii) tissue-specific demethylation associated with the onset of expression of the gene.
我们在此报告一项对单拷贝组织特异性基因特定位点甲基化状态发育变化的详细研究,研究对象涵盖从生殖细胞到早期胚胎再到成体组织。小鼠载脂蛋白AI基因5'端的两个位点在排卵后的未受精卵母细胞中未甲基化,而在精子中甲基化。相反,位于该基因上游的第三个位点在卵母细胞和精子中均甲基化,而基因内的一个CpG岛未甲基化。无论来自母方还是父方,甲基化位点在早期胚胎(8 - 16细胞期)经历去甲基化,并在囊胚晚期一直保持未甲基化状态。在原肠胚形成过程中,非CpG岛位点发生甲基化,随后在与该基因表达平行的组织(肝脏和肠道)中的特定位点逐渐去甲基化。因此,载脂蛋白AI基因成熟的组织特异性甲基化模式的形成涉及以下三个主要事件:(i)生殖细胞甲基化模式的消除(在8至16细胞期),(ii)通过非CpG岛位点的从头甲基化形成新的甲基化模式(在原肠胚形成过程中),以及(iii)与基因表达起始相关的组织特异性去甲基化。