Efficient differentiation of murine embryonic stem cells requires the binding of CXXC finger protein 1 to DNA or methylated histone H3-Lys4
Efficient differentiation of murine embryonic stem cells requires the binding of CXXC finger protein 1 to DNA or methylated histone H3-Lys4
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DOI：
10.1016/j.gene.2016.08.048
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发表时间：
2016-12-05
期刊：
影响因子：
3.5
通讯作者：
Skalnik, David G.
中科院分区：
文献类型：
--
作者：
Mahadevan, Jyothi;Skalnik, David G.
哺乳动物 CXXC 指蛋白 1 (Cfp1) 是一种 DNA 结合蛋白，是 Setd1 组蛋白甲基转移酶复合物的组成部分，也是组蛋白和胞嘧啶甲基化的关键表观遗传调节因子。缺乏 Cfp1 的小鼠胚胎干 (ES) 细胞在许多 CpG 岛表现出组蛋白 H3-Lys4 三甲基化 (H3K4me3) 的缺失，并且该表观遗传标记错误定位到异染色亚核结构域。此外，这些细胞无法在体外进行细胞分化。引入 Cfp1 表达载体后，这些缺陷得以修复。 Cfp1 包含 N 端植物同源结构域 (PHD)，这是在染色质相关蛋白中经常观察到的基序，充当组蛋白标记的读取模块。在这里，我们报告 Cfp1 PHD 结构域直接且特异性地与组蛋白 H3K4mel/me2/me3 标记结合。在关键 Cfp1 PHD 残基（Y28、D44 或 W49）处引入个体突变可在体外和体内消除这种组蛋白相互作用。 W49A 点突变不影响 Cfp1 挽救组蛋白 H3K4me3 对常染色质亚核结构域的适当限制的能力，也不影响 Cfp1 缺失 ES 细胞中的体外细胞分化的能力。同样，缺乏 DNA 结合活性的 Cfp1 突变形式 (C169A) 可以挽救体外细胞分化。然而，用 Cfp1 双突变体形式（W49A、C169A）拯救 Cfp1 缺失 ES 细胞会导致体外分化部分缺陷。这些数据将 Cfp1 PHD 结构域定义为组蛋白 H3K4me 标记的读取器，并提供证据表明该活性参与 ES 细胞中谱系定型的调节。 (C) 2016 Elsevier B.V. 保留所有权利。
Mammalian CXXC finger protein 1 (Cfp1) is a DNA-binding protein that is a component of the Setd1 histone methyltransferase complexes and is a critical epigenetic regulator of both histone and cytosine methylation. Murine embryonic stem (ES) cells lacking Cfp1 exhibit a loss of histone H3-Lys4 tri-methylation (H3K4me3) at many CpG islands, and a mis-localization of this epigenetic mark to heterochromatic sub-nuclear domains. Furthermore, these cells fail to undergo cellular differentiation in vitro. These defects are rescued upon introduction of a Cfp1-expression vector. Cfp1 contains an N-terminal plant homeodomain (PHD), a motif frequently observed in chromatin associated proteins that functions as a reader module of histone marks. Here, we report that the Cfp1 PHD domain directly and specifically binds to histone H3K4mel/me2/me3 marks. Introduction of individual mutations at key Cfp1 PHD residues (Y28, D44, or W49) ablates this histone interaction both in vitro and in vivo. The W49A point mutation does not affect the ability of Cfp1 to rescue appropriate restriction of histone H3K4me3 to euchromatic sub-nuclear domains or in vitro cellular differentiation in Cfp1-null ES cells. Similarly, a mutated form of Cfp1 that lacks DNA-binding activity (C169A) rescues in vitro cellular differentiation. However, rescue of Cfp1-null ES cells with a double mutant form of Cfp1 (W49A, C169A) results in partially defective in vitro differentiation. These data define the Cfp1 PHD domain as a reader of histone H3K4me marks and provide evidence that this activity is involved in the regulation of lineage commitment in ES cells. (C) 2016 Elsevier B.V. All rights reserved.