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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DOI：
10.1016/j.molcel.2015.05.017
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发表时间：
2015-07-02
期刊：
影响因子：
16
通讯作者：
Allis, C. David
中科院分区：
文献类型：
--
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
组蛋白修饰和 DNA 甲基化与不同的表观遗传“景观”相关，但两者之间的详细机制和功能联系仍不清楚。使用 DNA 甲基转移酶 Dnmt3a 的 ATRX-DNMT3-DNMT3L (ADD) 结构域作为范例，我们应用蛋白质工程来剖析将该酶招募到小鼠胚胎干细胞 (ESC) 染色质特定区域的分子相互作用。通过使 ADD 结构域对组蛋白修饰（特别是 H3K4 甲基化或 H3T3 磷酸化）不敏感，我们证明了 Dnmt3a 结合和活性失调的后果。 Dnmt3a 突变体靶向 H3K4me3 启动子会降低发育基因子集中的基因表达并改变 ESC 分化，而另一个突变体在有丝分裂过程中与 H3T3ph 的异常结合会促进染色体不稳定。我们的研究支持了这样的普遍观点，即组蛋白修饰“读取”和 DNA 甲基化在哺乳动物细胞中密切相关，并为染色质相关蛋白的功能评估提供了一条途径。
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.
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影响因子：
64.5
作者：
Gifford CA;Ziller MJ;Gu H;Trapnell C;Donaghey J;Tsankov A;Shalek AK;Kelley DR;Shishkin AA;Issner R;Zhang X;Coyne M;Fostel JL;Holmes L;Meldrim J;Guttman M;Epstein C;Park H;Kohlbacher O;Rinn J;Gnirke A;Lander ES;Bernstein BE;Meissner A
通讯作者：
Meissner A
影响因子：
64.5
作者：
Guenther, Matthew G.;Levine, Stuart S.;Young, Richard A.
通讯作者：
Young, Richard A.
影响因子：
9.2
作者：
Lehnertz, B;Ueda, Y;Peters, AHFM
通讯作者：
Peters, AHFM
染色质结合模块的组合分析揭示了多位点歧视。
影响因子：
14.8
作者：
Garske, Adam L.;Oliver, Samuel S.;Wagner, Elise K.;Musselman, Catherine A.;LeRoy, Gary;Garcia, Benjamin A.;Kutateladze, Tatiana G.;Denu, John M.
通讯作者：
Denu, John M.
影响因子：
30.8
作者：
Jones, PL;Veenstra, GJC;Wolffe, AP
通讯作者：
Wolffe, AP