Loss of Polycomb Group Protein Pcgf1 Severely Compromises Proper Differentiation of Embryonic Stem Cells
Loss of Polycomb Group Protein Pcgf1 Severely Compromises Proper Differentiation of Embryonic Stem Cells
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多梳族蛋白 Pcgf1 的丢失严重影响胚胎干细胞的正常分化
DOI：
10.1038/srep46276
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发表时间：
2017-04
影响因子：
4.6
通讯作者：
Qin Jinzhong
中科院分区：
文献类型：
--
作者：
Yan Yun;Zhao Wukui;Huang Yikai;Tong Huan;Xia Yin;Jiang Qing;Qin Jinzhong
Polycomb 抑制复合物 1 (PRC1) 对于胚胎干 (ES) 细胞的命运决定至关重要。新出现的证据表明，PRC1 复合体的六种主要变体（由相互排斥的 Pcgf 亚基定义）调节着不同的生物过程，但人们对每种版本的 PRC1 指导和维持细胞命运的机制知之甚少。在这里，我们通过 CRISPR/Cas9 技术破坏了小鼠 ES 细胞中的 Pcgf1（也称为 Nspc1，是六个 Pcgf 旁系同源物之一）。我们发现，尽管这些突变细胞具有活力并保留正常的自我更新，但它们在体外分化中表现出严重的缺陷。为了更好地了解 Pcgf1 在分化转录控制中的作用，我们使用 RNA-seq 分析了 Pcgf1 缺陷细胞的 mRNA 图谱。有趣的是，我们发现 Pcgf1 正向调节参与外胚层和中胚层分化的必需转录因子的表达，揭示了 Pcgf1 在 ES 细胞谱系规范过程中基因激活中的意外功能。染色质免疫沉淀实验表明，Pcgf1 缺失导致 Ring1B 及其相关的 H2AK119ub1 标记与靶基因的结合减少。总而言之，我们的结果表明 Pcgf1 在 ES 细胞维持期间的基因激活中具有意想不到的功能。
The Polycomb repressive complex 1 (PRC1) is essential for fate decisions of embryonic stem (ES) cells. Emerging evidence suggests that six major variants of PRC1 complex, defined by the mutually exclusive presence of Pcgf subunit, regulate distinct biological processes, yet very little is known about the mechanism by which each version of PRC1 instructs and maintains cell fate. Here, we disrupted the Pcgf1, also known as Nspc1 and one of six Pcgf paralogs, in mouse ES cells by the CRISPR/Cas9 technology. We showed that although these mutant cells were viable and retained normal self-renewal, they displayed severe defects in differentiationin vitro. To gain a better understanding of the role of Pcgf1 in transcriptional control of differentiation, we analysed mRNA profiles from Pcgf1 deficient cells using RNA-seq. Interestingly, we found that Pcgf1 positively regulated expression of essential transcription factors involved in ectoderm and mesoderm differentiation, revealing an unexpected function of Pcgf1 in gene activation during ES cell lineage specification. Chromatin immunoprecipitation experiments demonstrated that Pcgf1 deletion caused a decrease in Ring1B and its associated H2AK119ub1 mark binding to target genes. Altogether, our results suggested an unexpected function of Pcgf1 in gene activation during ES cell maintenance.
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影响因子：
7.7
作者：
Farcas AM;Blackledge NP;Sudbery I;Long HK;McGouran JF;Rose NR;Lee S;Sims D;Cerase A;Sheahan TW;Koseki H;Brockdorff N;Ponting CP;Kessler BM;Klose RJ
通讯作者：
Klose RJ
影响因子：
23.9
作者：
Qin, Jinzhong;Whyte, Warren A.;Anderssen, Endre;Apostolou, Effie;Chen, Hsu-Hsin;Akbarian, Schahram;Bronson, Roderick T.;Hochedlinger, Konrad;Ramaswamy, Sridhar;Young, Richard A.;Hock, Hanno
通讯作者：
Hock, Hanno
影响因子：
8.8
作者：
Boles, Nathan C.;Hirsch, Sarah E.;Fasano, Christopher A.
通讯作者：
Fasano, Christopher A.
影响因子：
64.5
作者：
Czermin, B;Melfi, R;Pirrotta, V
通讯作者：
Pirrotta, V
影响因子：
10.5
作者：
Lagarou, Anna;Mohd-Sarip, Adone;Verrijzer, C. Peter
通讯作者：
Verrijzer, C. Peter