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首页> 外文期刊>Molecular and Cellular Biology >Parent-of-Origin-Specific Binding of Nuclear Hormone Receptor Complexes in the H19-Igf2 Imprinting Control Region
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Parent-of-Origin-Specific Binding of Nuclear Hormone Receptor Complexes in the H19-Igf2 Imprinting Control Region

机译：H19-Igf2印迹控制区域中核激素受体复合物的原产地特异性结合。

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Parent-of-origin-specific expression of the mouse insulin-like growth factor 2 gene (Igf2) and the closely linked H19 gene located on distal chromosome 7 is regulated by a 2.4-kb imprinting control region (ICR) located upstream of the H19 gene. In somatic cells, the maternally and paternally derived ICRs are hypo- and hypermethylated, respectively, with the former binding the insulator protein CCCTC-binding factor (CTCF) and acting to block access of enhancers to the Igf2 promoter. Here we report on a detailed in vivo footprinting analysis—using ligation-mediated PCR combined with in vivo dimethyl sulfate, DNase I, or UV treatment—of ICR sequences located outside of the CTCF binding domains. In mouse primary embryo fibroblasts carrying only maternal or paternal copies of distal chromosome 7, we have identified five prominent footprints specific to the maternal ICR. Each of the five footprinted areas contains at least two nuclear hormone receptor hexad binding sites arranged with irregular spacing. When combined with fibroblast nuclear extracts, these sequences interact with complexes containing retinoic X receptor alpha and estrogen receptor beta. More significantly, the footprint sequences bind nuclear hormone receptor complexes in male, but not female, germ cell extracts purified from fetuses at a developmental stage corresponding to the time of establishment of differential ICR methylation. These data are consistent with the possibility that nuclear hormone receptor complexes participate in the establishment of differential ICR methylation imprinting in the germ line.
机译：小鼠胰岛素样生长因子2基因（ Igf2 ）和位于远端7号染色体上的紧密连接的 H19 基因的原代亲本特异性表达受a调控。位于 H19 基因上游的2.4 kb印迹控制区（ICR）。在体细胞中，母本和父本来源的ICR分别被低甲基化和超甲基化，前者结合绝缘蛋白CCCTC结合因子（CTCF）并阻止增强子进入 Igf2 启动子。在这里，我们对位于CTCF结合域之外的ICR序列进行了详细的体内足迹分析（使用连接介导的PCR结合体内硫酸二甲酯，DNase I或UV处理）。在仅携带母本或父本拷贝的第7号远端染色体的小鼠原代成纤维细胞中，我们鉴定出了五个母本ICR特有的显着印迹。五个足迹区域的每一个都包含至少两个以不规则间距排列的核激素受体六聚体结合位点。当与成纤维细胞核提取物结合时，这些序列与含有视黄酸X受体α和雌激素受体β的复合物相互作用。更重要的是，足迹序列结合了从胎儿纯化的雄性而不是雌性的生殖细胞提取物中的核激素受体复合物，该发育阶段对应于建立差异ICR甲基化的时间。这些数据与核激素受体复合物参与种系中差异性ICR甲基化印迹建立的可能性一致。

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