Regulation of mouse embryonic stem cell self-renewal by a Yes-YAP-TEAD2 signaling pathway downstream of LIF
Regulation of mouse embryonic stem cell self-renewal by a Yes-YAP-TEAD2 signaling pathway downstream of LIF
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DOI：
10.1242/jcs.075796
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发表时间：
2011-04-01
影响因子：
4
通讯作者：
Anneren, Cecilia
中科院分区：
文献类型：
--
作者：
Tamm, Christoffer;Bower, Nathalie;Anneren, Cecilia
细胞质酪氨酸激酶Yes先前已被证明通过白血病抑制因子（LIF）和血清中的一种或多种因子的未知下游途径在维持小鼠和人胚胎干（ES）自我更新中具有重要作用。在这里，我们表明，TEAD 2和它的转录辅激活因子，是相关的蛋白雅普，合作的信号通路下游的是。我们发现，雅普，TEAD 2和Yes在自我更新的ES细胞中高度表达，被LIF和血清激活，并在细胞被诱导分化时下调。我们还证明，激酶活性的是结合和磷酸化雅普，并激活YAP-TEAD 2依赖的转录。我们发现TEAD 2与Oct-3/4启动子直接相关。此外，Yes途径的激活诱导Oct-3/4和Nanog启动子的活性，而抑制该途径则抑制启动子活性。Nanog反过来抑制TEAD 2依赖性启动子活性，而siRNA介导的Nanog敲低诱导了它，这表明了一个负调控反馈回路。用抑制性TEAD 2-EnR的细胞的附加体超转染诱导内胚层分化，这表明该途径对于ES细胞的维持是必需的。
The cytoplasmic tyrosine kinase Yes has previously been shown to have an important role in maintaining mouse and human embryonic stem (ES) self-renewal through an unknown pathway downstream of leukemia inhibitory factor (LIF) and one or more factors in serum. Here, we show that TEAD2 and its transcriptional co-activator, the Yes-associated protein YAP, co-operate in a signaling pathway downstream of Yes. We show that YAP, TEAD2 and Yes are highly expressed in self-renewing ES cells, are activated by LIF and serum, and are downregulated when cells are induced to differentiate. We also demonstrate that kinase-active Yes binds and phosphorylates YAP, and activates YAP-TEAD2-dependent transcription. We found that TEAD2 associates directly with the Oct-3/4 promoter. Moreover, activation of the Yes pathway induced activity of the Oct-3/4 and Nanog promoters, whereas suppression of this pathway inhibited promoter activity. Nanog, in turn, suppressed TEAD2-dependent promoter activity, whereas siRNA-mediated knockdown of Nanog induced it, suggesting a negative regulatory feedback loop. Episomal supertransfection of cells with inhibitory TEAD2-EnR induced endodermal differentiation, which suggests that this pathway is necessary for ES cell maintenance.