Self-renewal of rodent embryonic stem cells is enhanced by partial inhibition of glycogen synthase kinase-3 (Gsk3; refs 1, 2). This effect has variously been attributed to stimulation of Wnt signalling by β-catenin1, stabilization of Myc protein3 and global de-inhibition of anabolic processes4. Here we demonstrate that β-catenin is not necessary for embryonic stem cell identity or expansion, but its absence eliminates the self-renewal response to Gsk3 inhibition. Responsiveness is fully restored by truncated β-catenin lacking the carboxy-terminal transactivation domain5. However, requirement for Gsk3 inhibition is dictated by expression of T-cell factor 3 (Tcf3) and mediated by direct interaction with β-catenin. Tcf3 localizes to many pluripotency genes6 in embryonic stem cells. Our findings confirm that Tcf3 acts as a transcriptional repressor and reveal that β-catenin directly abrogates Tcf3 function. We conclude that Gsk3 inhibition stabilizes the embryonic stem cell state primarily by reducing repressive influence on the core pluripotency network.
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We thank N. Lyashenko and C. Hartmann for discussion and exchange of unpublished data. We also thank A. Martinez-Arias for comments on the manuscript. We are grateful to B. Merrill (University of Illinois at Chicago, USA) for generously providing Tcf3 mutant embryonic stem cells and to B. Doble (Stem Cell and Cancer Research Institute, McMaster University, Canada) and J. Woodgett (Samuel Lunenfeld Research Institute, Mount Sinai Hospital and University of Toronto, Canada) for Gsk3 mutant embryonic stem cells. We thank R. Grosschedl for the β-catenin ΔC construct. Gsk3 inhibitors, compounds A–G, were provided by Pfizer. R. Walker and P. Humphreys supported flow cytometry and imaging, respectively. The study was financially supported by the Biotechnology and Biological Sciences Research Council and the Medical Research Council of the United Kingdom, the Wellcome Trust and the European Commission FP7 project EuroSyStem. S.G-L. was supported by a CONACYT Studentship. A.S. is a Medical Research Council Professor.
The authors declare no competing financial interests.
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Wray, J., Kalkan, T., Gomez-Lopez, S. et al. Inhibition of glycogen synthase kinase-3 alleviates Tcf3 repression of the pluripotency network and increases embryonic stem cell resistance to differentiation. Nat Cell Biol 13, 838–845 (2011). https://doi.org/10.1038/ncb2267
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