De novo DNA methyltransferase is essential for self-renewal, but not for differentiation, in hematopoietic stem cells.
De novo DNA methyltransferase is essential for self-renewal, but not for differentiation, in hematopoietic stem cells.
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DOI：
10.1084/jem.20060750
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发表时间：
2007-04-16
期刊：
影响因子：
--
通讯作者：
Nakauchi H
中科院分区：
文献类型：
--
作者：
Tadokoro Y;Ema H;Okano M;Li E;Nakauchi H
DNA甲基化是发育所必需的表观遗传修饰。DNA甲基转移酶DNMT3a和Dnmt3b在原肠胚胎和分化生殖细胞中执行从头DNA甲基化。人们一直认为这些酶通常在调节细胞分化方面发挥作用。为了验证这一假设，我们研究了DNMT3A和DNMT3B在成人干细胞中的作用。CD34DNMT3A/Low、c-−+、Sca-1+、谱系标志−(CD34DNMT3A/Low、c-Kit+、Sca-1+)细胞是高度富含造血干细胞(HSC)的小鼠骨髓细胞的一部分，表达−3a和Dnmt3b。通过逆转录病毒Cre基因转导，我们有条件地干扰了CD34LKSL细胞中的DNMT3A、DNMT3b或DNMT3A和DNMT3b(DNMT3A/DNMT3b)，这些基因的功能区两侧有两个loxP位点。我们发现DNMT3A和DNMT3B在造血细胞分化过程中起着从头DNA甲基转移酶的作用。出乎意料的是，体外集落分析和体内移植试验表明，在DNMT3A-、Dnmt3b-和DNMT3A/Dnmt3b缺陷的HSC中，髓系和淋巴系的分化潜能都保持不变。然而，在移植试验中，DNMT3A/Dnmt3b缺陷的HSCs，而不是DNMT3A或Dnmt3b缺陷的HSCs，不能进行长期重建。这些发现确立了DNMT3A和DNMT3B的DNA甲基化在HSC自我更新中的关键作用。
DNA methylation is an epigenetic modification essential for development. The DNA methyltransferases Dnmt3a and Dnmt3b execute de novo DNA methylation in gastrulating embryos and differentiating germline cells. It has been assumed that these enzymes generally play a role in regulating cell differentiation. To test this hypothesis, we examined the role of Dnmt3a and Dnmt3b in adult stem cells. CD34−/low, c-Kit+, Sca-1+, lineage marker− (CD34− KSL) cells, a fraction of mouse bone marrow cells highly enriched in hematopoietic stem cells (HSCs), expressed both Dnmt3a and Dnmt3b. Using retroviral Cre gene transduction, we conditionally disrupted Dnmt3a, Dnmt3b, or both Dnmt3a and Dnmt3b (Dnmt3a/Dnmt3b) in CD34− KSL cells purified from mice in which the functional domains of these genes are flanked by two loxP sites. We found that Dnmt3a and Dnmt3b function as de novo DNA methyltransferases during differentiation of hematopoietic cells. Unexpectedly, in vitro colony assays and in vivo transplantation assays showed that both myeloid and lymphoid lineage differentiation potentials were maintained in Dnmt3a-, Dnmt3b-, and Dnmt3a/Dnmt3b-deficient HSCs. However, Dnmt3a/Dnmt3b-deficient HSCs, but not Dnmt3a- or Dnmt3b-deficient HSCs, were incapable of long-term reconstitution in transplantation assays. These findings establish a critical role for DNA methylation by Dnmt3a and Dnmt3b in HSC self-renewal.
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影响因子：
64.5
作者：
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