Regulation of mouse embryonic stem cell self-renewal by a Yes–YAP–TEAD2 signaling pathway downstream of LIF

Abstract

SummaryThe 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.

The Hippo pathway regulates stem cell proliferation, self-renewal, and differentiation

“…However, the mechanism responsible for specificity of YAP and TAZ in BMP and TGFbeta signaling is not known. Another report suggests the regulation of YAP by LIF, which is required by mouse but not human ESCs, which is another possible explanation for the role of YAP and TAZ in the self-renewal of human and mouse ESCs (Tamm et al, 2011).…”

“…However, only until recently, it was demonstrated that YAP as well as its transcription factor partner TEAD promotes ES cell self-renewal. When mouse ESCs were induced to differentiate, YAP was inactivated as indicated by decreased protein level and increased phosphorylation (Lian et al, 2010;Tamm et al, 2011). However, YAP overexpression prevents mouse ESCs from differentiation even under differentiation condition such as leukemia inhibitory factor (LIF) withdrawal (Lian et al, 2010;Tamm et al, 2011).…”

“…When mouse ESCs were induced to differentiate, YAP was inactivated as indicated by decreased protein level and increased phosphorylation (Lian et al, 2010;Tamm et al, 2011). However, YAP overexpression prevents mouse ESCs from differentiation even under differentiation condition such as leukemia inhibitory factor (LIF) withdrawal (Lian et al, 2010;Tamm et al, 2011). In addition, YAP knockdown leads to loss of pluripotency under a condition that would normally support stemness (Lian et al, 2010;Tamm et al, 2011).…”

“…However, YAP overexpression prevents mouse ESCs from differentiation even under differentiation condition such as leukemia inhibitory factor (LIF) withdrawal (Lian et al, 2010;Tamm et al, 2011). In addition, YAP knockdown leads to loss of pluripotency under a condition that would normally support stemness (Lian et al, 2010;Tamm et al, 2011). Chromatin immunoprecipitation (ChIP) experiments demonstrated that YAP-TEAD bind to promoters of many stemnesspromoting genes such as Oct4 (Lian et al, 2010;Tamm et al, 2011).…”

“…In addition, YAP knockdown leads to loss of pluripotency under a condition that would normally support stemness (Lian et al, 2010;Tamm et al, 2011). Chromatin immunoprecipitation (ChIP) experiments demonstrated that YAP-TEAD bind to promoters of many stemnesspromoting genes such as Oct4 (Lian et al, 2010;Tamm et al, 2011). Intriguingly, knockdown of TAZ in human ESCs also led to differentiation and loss of pluripotency, although YAP was intact (Varelas et al, 2008).…”