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Imprinted DNA methylation reconstituted at a non-imprinted locus

机译:印迹DNA甲基化在非印迹位点重建

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Background In mammals, tight regulation of cytosine methylation is required for embryonic development and cellular differentiation. The trans -acting DNA methyltransferases that catalyze this modification have been identified and characterized; however, these proteins lack sequence specificity, leaving the mechanism of targeting unknown. A cis -acting regulator within the Rasgrf1 imprinting control region (ICR) is necessary for establishment and maintenance of local imprinted methylation. Here, we investigate whether 3-kb of sequence from the Rasgrf1 ICR is sufficient to direct appropriate imprinted methylation and target gene expression patterns when ectopically inserted at the Wnt1 locus. Results The Rasgrf1 ICR at Wnt1 lacked somatic methylation when maternally transmitted and was fully methylated upon paternal transmission, consistent with its behavior at the Rasgrf1 locus. It was unmethylated in the female germline and was enriched for methylation in the male germline, though not to the levels seen at the endogenous Rasgrf1 allele. Wnt1 expression was not imprinted by the ectopic ICR, likely due to additional sequences being required for this function. Conclusions We have identified sequences that are sufficient for partial establishment and full maintenance of the imprinted DNA methylation patterns. Because full somatic methylation can occur without full gametic methylation, we infer that somatic methylation of the Rasgrf1 ICR is not simply a consequence of maintained gametic methylation.
机译:背景技术在哺乳动物中,需要严格调节胞嘧啶甲基化以用于胚胎发育和细胞分化。已经鉴定和表征了催化这种修饰的反式DNA甲基转移酶。然而,这些蛋白质缺乏序列特异性,使得靶向机制尚不清楚。 Rasgrf1印迹控制区域(ICR)内的顺式调节剂对于建立和维持局部印迹甲基化是必要的。在这里,我们调查从Rasgrf1 ICR的3 kb序列是否足以在异位插入Wnt1基因座时指导适当的印迹甲基化和靶基因表达模式。结果Wnt1的Rasgrf1 ICR在母体传播时缺乏体细胞甲基化,而在父本传播时被完全甲基化,这与其在Rasgrf1基因座上的行为一致。它在雌性种系中未甲基化,而在雄性种系中富含甲基化,尽管没有达到内源Rasgrf1等位基因所见的水平。 Wnt1表达未被异位ICR标记,可能是由于此功能需要其他序列。结论我们确定了足以部分构建和完全维持印迹DNA甲基化模式的序列。因为完全的体细胞甲基化可以发生而没有完全的配子体甲基化,所以我们推断Rasgrf1 ICR的体细胞甲基化不仅仅是保持配子体甲基化的结果。

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