CpG 位点特异性羟甲基胞嘧啶向甲基胞嘧啶的转化超越了 DNA 复制。

CpG site-specific alteration of hydroxymethylcytosine to methylcytosine beyond DNA replication.

机构信息

RIKEN Omics Science Center, 1-7-22 Suehiro-cho, Yokohama, Kanagawa 230-0045, Japan.

出版信息

Biochem Biophys Res Commun. 2012 Sep 14;426(1):141-7. doi: 10.1016/j.bbrc.2012.08.053. Epub 2012 Aug 17.

Abstract

Hydroxymethylcytosines (hmC), one of several reported cytosine modifications, was recently found to be enriched in embryonic stem cells and neuronal cells, and thought to play an important role in regulating gene expression and cell specification. However, unlike methylcytosines (mC), the fate of hmC beyond DNA replication is not well understood. Here, to monitor the status of hmC during DNA replication, we prepared a stable episomal vector-based monitoring system called MoCEV in 293T cells. The MoCEV system containing fully hydroxymethylated-cytosine fragments revealed a significant modification towards mC after several rounds of DNA replication. Strikingly this modification was specifically observed at the CpG sites (71.9% of cytosines), whereas only 1.1% of modified cytosines were detected at the non-CpG sites. Since the unmodified MoCEV did not undergo any DNA methylation during cell division, the results strongly suggest that somatic cells undergo hmC to mC specifically at the CpG sites during cell division.

摘要

羟甲基胞嘧啶(hmC)是几种已报道的胞嘧啶修饰之一,最近发现在胚胎干细胞和神经元细胞中富集,并被认为在调节基因表达和细胞特化中发挥重要作用。然而,与甲基胞嘧啶(mC)不同,hmC 在 DNA 复制后的命运尚不清楚。在这里,为了在 DNA 复制过程中监测 hmC 的状态,我们在 293T 细胞中制备了一种稳定的基于附加体的监测系统,称为 MoCEV。MoCEV 系统包含完全羟甲基化的胞嘧啶片段,在经过几轮 DNA 复制后显示出向 mC 的显著修饰。引人注目的是,这种修饰专门发生在 CpG 位点(71.9%的胞嘧啶),而在非 CpG 位点仅检测到 1.1%的修饰胞嘧啶。由于未修饰的 MoCEV 在细胞分裂过程中没有发生任何 DNA 甲基化,因此结果强烈表明体细胞在细胞分裂过程中特异性地将 hmC 转化为 CpG 位点的 mC。

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