Inhibitory action of an ERK1/2 inhibitor on primitive endoderm cell differentiation from mouse embryonic stem cells.
Inhibitory action of an ERK1/2 inhibitor on primitive endoderm cell differentiation from mouse embryonic stem cells.
复制标题
ERK1/2 抑制剂对小鼠胚胎干细胞原始内胚层细胞分化的抑制作用。
DOI：
10.1016/j.bbrc.2019.03.081
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
and K. Kitajima.
中科院分区：
文献类型：
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作者：
H. Tabata;T. Hara;and K. Kitajima.
细胞外信号调节激酶1/2（ERK 1/2）和糖原合成酶激酶3β（GSK 3 β）抑制剂的组合（称为2 i）被广泛用于维持小鼠胚胎干细胞（ESC）的体外多能性。在没有2 i的情况下，一些小鼠ESC自发地产生原始内胚层（PrE）细胞，而2 i完全阻断这种PrE细胞分化。然而，2 i对PrE细胞分化的抑制作用的分子机制仍不清楚。通过转录因子Gata 4的强制表达实现稳健的PrE细胞诱导。在这里，我们分析了2 i如何抑制PrE细胞分化使用小鼠胚胎干细胞携带诱导型Gata 4表达盒。我们发现2 i有效地抑制Gata 4诱导的PrE细胞分化，ERK 1/2抑制剂负责这种作用。我们进一步揭示了Gata 4的转录激活能力是PrE细胞诱导和ERK 1/2抑制剂对其破坏所必需的。Gata 4蛋白的Ser 105、Ser 266和Ser 411的磷酸化不参与PrE细胞的诱导。过表达ERK 1/2底物Klf 4抑制Gata 4介导的转录激活。我们的数据表明，ERK 1/2通过Gata 4的间接转录激活支持PrE细胞诱导。
A combination of extracellular signal-regulated kinase 1/2 (ERK1/2) and glycogen synthase kinase 3β (GSK3β) inhibitors, called 2i, is widely used for maintaining the pluripotency of mouse embryonic stem cells (ESCs)in vitro. Without 2i, a few mouse ESCs spontaneously gives rise to primitive endoderm (PrE) cells, whereas 2i completely blocks this PrE cell differentiation. However, the molecular mechanisms underlying the inhibitory action of 2i on PrE cell differentiation remain unclear. Robust PrE cell induction is achieved by enforced expression of the transcription factor Gata4. Here, we analyzed how 2i inhibits the PrE cell differentiation using mouse ESCs carrying an inducible Gata4 expression cassette. We found that 2i effectively inhibited the Gata4-induced PrE cell differentiation and the ERK1/2 inhibitor was responsible for this effect. We further revealed that the transcriptional activation ability of Gata4 was necessary for PrE cell induction and its disruption by the ERK1/2 inhibitor. The phosphorylation of Ser105, Ser266, and Ser411of the Gata4 protein was not involved in the PrE cell induction. Overexpression of Klf4, an ERK1/2 substrate, inhibited the Gata4-mediated transcriptional activation. Our data indicated that ERK1/2 supported the PrE cell induction via the indirect transcriptional activation of Gata4.