Engineering of a Histone-Recognition Domain in Dnmt3a Alters the Epigenetic Landscape and Phenotypic Features of Mouse ESCs.
Engineering of a Histone-Recognition Domain in Dnmt3a Alters the Epigenetic Landscape and Phenotypic Features of Mouse ESCs.
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Dnmt3a 中组蛋白识别域的工程改变了小鼠 ESC 的表观遗传景观和表型特征。
DOI：
10.1016/j.molcel.2018.01.014
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发表时间：
2018-02-01
期刊：
影响因子：
16
通讯作者：
Allis CD
中科院分区：
文献类型：
--
作者：
Noh KM;Wang H;Kim HR;Wenderski W;Fang F;Li CH;Dewell S;Hughes SH;Melnick AM;Patel DJ;Li H;Allis CD
组蛋白修饰和DNA甲基化与不同的表观遗传“景观”有关，但两者之间详细的机制和功能联系仍不清楚。使用的ATRX-DNMT 3-DNMT 3L（ADD）域的DNA甲基转移酶Dnmt 3a作为范例，我们应用蛋白质工程解剖的分子相互作用的基础上，这种酶在小鼠胚胎干细胞（ESC）的特定区域的染色质的招聘。通过使ADD结构域对组蛋白修饰，特别是H3 K4甲基化或H3 T3磷酸化不敏感，我们证明了Dnmt 3a结合和活性失调的后果。Dnmt 3a突变体靶向H3 K4 me 3启动子降低了发育基因亚组中的基因表达并改变了ESC分化，而另一突变体在有丝分裂期间与H3 T3 ph的异常结合促进了染色体不稳定性。我们的研究支持了组蛋白修饰“阅读”和DNA甲基化在哺乳动物细胞中密切相关的普遍观点，并为染色质相关蛋白的功能评估提供了一种途径。
Histone modification and DNA methylation are associated with varying epigenetic "landscapes," but detailed mechanistic and functional links between the two remain unclear. Using the ATRX-DNMT3-DNMT3L (ADD) domain of the DNA methyltransferase Dnmt3a as a paradigm, we apply protein engineering to dissect the molecular interactions underlying the recruitment of this enzyme to specific regions of chromatin in mouse embryonic stem cells (ESCs). By rendering the ADD domain insensitive to histone modification, specifically H3K4 methylation or H3T3 phosphorylation, we demonstrate the consequence of dysregulated Dnmt3a binding and activity. Targeting of a Dnmt3a mutant to H3K4me3 promoters decreases gene expression in a subset of developmental genes and alters ESC differentiation, whereas aberrant binding of another mutant to H3T3ph during mitosis promotes chromosome instability. Our studies support the general view that histone modification "reading" and DNA methylation are closely coupled in mammalian cells, and suggest an avenue for the functional assessment of chromatin-associated proteins.
影响因子：
16
作者：
Noh, Kyung-Min;Wang, Haibo;Kim, Hyunjae R.;Wenderski, Wendy;Fang, Fang;Li, Charles H.;Dewell, Scott;Hughes, Stephen H.;Melnick, Ari M.;Patel, Dinshaw J.;Li, Haitao;Allis, C. David
通讯作者：
Allis, C. David