Abstract
The co-occupancy of Tcf3 with Oct4, Sox2 and Nanog on embryonic stem cell (ESC) chromatin indicated that Tcf3 has been suggested to play an integral role in a poorly understood mechanism underlying Wnt-dependent stimulation of mouse ESC self-renewal of mouse ESCs. Although the conventional view of Tcf proteins as the β-catenin-binding effectors of Wnt signalling suggested Tcf3–β-catenin activation of target genes would stimulate self-renewal, here we show that an antagonistic relationship between Wnt3a and Tcf3 on gene expression regulates ESC self-renewal. Genetic ablation of Tcf3 replaced the requirement for exogenous Wnt3a or GSK3 inhibition for ESC self-renewal, demonstrating that inhibition of Tcf3 repressor is the necessary downstream effect of Wnt signalling. Interestingly, both Tcf3–β-catenin and Tcf1–β-catenin interactions contributed to Wnt stimulation of self-renewal and gene expression, and the combination of Tcf3 and Tcf1 recruited Wnt-stabilized β-catenin to Oct4 binding sites on ESC chromatin. This work elucidates the molecular link between the effects of Wnt and the regulation of the Oct4/Sox2/Nanog network.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Accession codes
References
Kim, J., Chu, J., Shen, X., Wang, J. & Orkin, S. H. An extended transcriptional network for pluripotency of embryonic stem cells. Cell 132, 1049–1061 (2008).
Liang, J. et al. Nanog and Oct4 associate with unique transcriptional repression complexes in embryonic stem cells. Nat. Cell Biol. 10, 731–739 (2008).
Boyer, L. A. et al. Core transcriptional regulatory circuitry in human embryonic stem cells. Cell 122, 947–956 (2005).
Loh, Y. H. et al. The Oct4 and Nanog transcription network regulates pluripotency in mouse embryonic stem cells. Nat. Genet. 38, 431–440 (2006).
Nichols, J. et al. Formation of pluripotent stem cells in the mammalian embryo depends on the POU transcription factor Oct4. Cell 95, 379–391 (1998).
Niwa, H., Miyazaki, J. & Smith, A. G. Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells. Nat. Genet. 24, 372–376 (2000).
Avilion, A. A. et al. Multipotent cell lineages in early mouse development depend on SOX2 function. Genes Dev. 17, 126–140 (2003).
Chambers, I. et al. Functional expression cloning of Nanog, a pluripotency sustaining factor in embryonic stem cells. Cell 113, 643–655 (2003).
Mitsui, K. et al. The homeoprotein Nanog is required for maintenance of pluripotency in mouse epiblast and ES cells. Cell 113, 631–642 (2003).
Marson, A. et al. Connecting microRNA genes to the core transcriptional regulatory circuitry of embryonic stem cells. Cell 134, 521–533 (2008).
Cole, M. F., Johnstone, S. E., Newman, J. J., Kagey, M. H. & Young, R. A. Tcf3 is an integral component of the core regulatory circuitry of embryonic stem cells. Genes Dev. 22, 746–755 (2008).
Tam, W. L. et al. T-cell factor 3 regulates embryonic stem cell pluripotency and self-renewal by the transcriptional control of multiple lineage pathways. Stem Cells 26, 2019–2031 (2008).
Yi, F., Pereira, L. & Merrill, B.J. Tcf3 functions as a steady state limiter of transcriptional programs of mouse embryonic stem cell self renewal. Stem Cells 26, 1951–1960 (2008).
MacDonald, B. T., Tamai, K. & He, X. Wnt/β-catenin signaling: components, mechanisms, and diseases. Dev. Cell 17, 9–26 (2009).
Brannon, M., Gomperts, M., Sumoy, L., Moon, R. T. & Kimelman, D. A β-catenin/XTcf-3 complex binds to the siamois promoter to regulate dorsal axis specification in Xenopus . Genes Dev. 11, 2359–2370 (1997).
Molenaar, M. et al. XTcf-3 transcription factor mediates β-catenin-induced axis formation in Xenopus embryos. Cell 86, 391–399 (1996).
van de Wetering, M. et al. Armadillo coactivates transcription driven by the product of the Drosophila segment polarity gene dTCF. Cell 88, 789–799 (1997).
Sato, N., Meijer, L., Skaltsounis, L., Greengard, P. & Brivanlou, A.H. Maintenance of pluripotency in human and mouse embryonic stem cells through activation of Wnt signaling by a pharmacological GSK-3-specific inhibitor. Nat. Med. 10, 55–63 (2004).
Doble, B. W., Patel, S., Wood, G. A., Kockeritz, L. K. & Woodgett, J. R. Functional redundancy of GSK-3α and GSK-3β in Wnt/β-catenin signaling shown by using an allelic series of embryonic stem cell lines. Dev Cell 12, 957–971 (2007).
Nichols, J., Silva, J., Roode, M. & Smith, A. Suppression of Erk signalling promotes ground state pluripotency in the mouse embryo. Development 136, 3215–3222 (2009).
Ying, Q. L. et al. The ground state of embryonic stem cell self-renewal. Nature 453, 519–523 (2008).
Kelly, K. F. et al. β-catenin enhances Oct-4 activity and reinforces pluripotency through a TCF-independent mechanism. Cell Stem Cell 8, 214–227 (2011).
Pereira, L., Yi, F. & Merrill, B. J. Repression of nanog gene transcription by tcf3 limits embryonic stem cell self-renewal. Mol. Cell Biol. 26, 7479–7491 (2006).
Nishiyama, A. et al. Uncovering early response of gene regulatory networks in ESCs by systematic induction of transcription factors. Cell Stem Cell 5, 420–433 (2009).
Raychaudhuri, S., Stuart, J. M. & Altman, R. B. Principal components analysis to summarize microarray experiments: application to sporulation time series. Pac. Symp. Biocomput. 2000, 455–466 (2000).
Arce, L., Yokoyama, N. N. & Waterman, M. L. Diversity of LEF/TCF action in development and disease. Oncogene 25, 7492–7504 (2006).
van de Wetering, M., Oosterwegel, M., Dooijes, D. & Clevers, H. Identification and cloning of TCF-1, a T lymphocyte-specific transcription factor containing a sequence-specific HMG box. EMBO J. 10, 123–132 (1991).
Galceran, J., Farinas, I., Depew, M. J., Clevers, H. & Grosschedl, R. Wnt3a-/- like phenotype and limb deficiency in Lef1(-/-)Tcf1(-/-) mice. Genes Dev. 13, 709–717 (1999).
Gregorieff, A., Grosschedl, R. & Clevers, H. Hindgut defects and transformation of the gastro-intestinal tract in Tcf4(-/-)/Tcf1(-/-) embryos. EMBO J. 23, 1825–1833 (2004).
Nguyen, H. et al. Tcf3 and Tcf4 are essential for long-term homeostasis of skin epithelia. Nat. Genet. 41, 1068–1075 (2009).
Biechele, T. L., Adams, A. M. & Moon, R. T. Transcription-based reporters of Wnt/β-catenin signaling. Cold Spring Harb. Protoc.doi:10.1101/pdb.prot5223 (2009).
Jho, E. H. et al. Wnt/β-catenin/Tcf signaling induces the transcription of Axin2, a negative regulator of the signaling pathway. Mol. Cell Biol. 22, 1172–1183 (2002).
Abu-Remaileh, M. et al. Oct-3/4 regulates stem cell identity and cell fate decisions by modulating Wnt/β-catenin signalling. EMBO J. 29, 3236–3248 (2010).
Takao, Y., Yokota, T. & Koide, H. β-catenin up-regulates Nanog expression through interaction with Oct-3/4 in embryonic stem cells. Biochem. Biophys. Res. Commun. 353, 699–705 (2007).
Wang, J. et al. A protein interaction network for pluripotency of embryonic stem cells. Nature 444, 364–368 (2006).
Acknowledgements
We thank K. Plath for helpful discussions and thoughtful comments on the manuscript, and M. Ko and A. Nishiyama for the inducible Tcf3 overexpression cell line. The work was financially supported by grants from the American Cancer Society (RSG GGC 112994, B.J.M.), the National Institutes of Health (R01-CA128571, B.J.M. and R01-GM065400, D.R.L.) and the Howard Hughes Medical Institute (D.R.L.).
Author information
Authors and Affiliations
Contributions
All authors designed and analysed experiments. F.Y., L.P., J.A.H., B.R.S. and C.M.Y. carried out experiments. F.Y., L.P. and B.J.M. wrote the manuscript. B.J.M. and D.R.L. supervised the project.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Information
Supplementary Information (PDF 868 kb)
Supplementary Information
Supplementary Table 1 (XLSX 161 kb)
Supplementary Information
Supplementary Table 2 (XLSX 139 kb)
Supplementary Information
Supplementary Table 3 (XLSX 59 kb)
Supplementary Information
Supplementary Table 4 (XLSX 10 kb)
Supplementary Information
Supplementary Table 5 (XLSX 12 kb)
Supplementary Information
Supplementary Table 6 (XLSX 10 kb)
Rights and permissions
About this article
Cite this article
Yi, F., Pereira, L., Hoffman, J. et al. Opposing effects of Tcf3 and Tcf1 control Wnt stimulation of embryonic stem cell self-renewal. Nat Cell Biol 13, 762–770 (2011). https://doi.org/10.1038/ncb2283
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/ncb2283
This article is cited by
-
JUNB affects hair follicle development and regeneration by promoting the proliferation of dermal papilla cells in goat
Chemical and Biological Technologies in Agriculture (2023)
-
The Wnt/TCF7L1 transcriptional repressor axis drives primitive endoderm formation by antagonizing naive and formative pluripotency
Nature Communications (2023)
-
Ascl1 and Ngn2 convert mouse embryonic stem cells to neurons via functionally distinct paths
Nature Communications (2023)
-
Sox2 levels regulate the chromatin occupancy of WNT mediators in epiblast progenitors responsible for vertebrate body formation
Nature Cell Biology (2022)
-
Integrated analysis reveals the alterations that LMNA interacts with euchromatin in LMNA mutation-associated dilated cardiomyopathy
Clinical Epigenetics (2021)