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Nanog and Oct4 associate with unique transcriptional repression complexes in embryonic stem cells

Nature Cell Biology volume 10, pages 731–739 (2008)Cite this article

Abstract

Nanog and Oct4 are essential transcription factors that regulate self-renewal and pluripotency of ES cells. However, the mechanisms by which Nanog and Oct4 modulate ES cell fate remain unknown. Through characterization of endogenous Nanog and Oct4 protein complexes in mouse ES cells, we found that these transcription factors interact with each other and associate with proteins from multiple repression complexes, including the NuRD, Sin3A and Pml complexes. In addition, Nanog, Oct4 and repressor proteins co-occupy Nanog-target genes in mouse ES cells, suggesting that Nanog and Oct4 together may communicate with distinct repression complexes to control gene transcription. To our surprise, of the various core components in the NuRD complex with which Nanog and Oct4 interact, Mta1 was preferred, whereas Mbd3 and Rbbp7 were either absent or present at sub-stoichiometric levels. We named this unique Hdac1/2- and Mta1/2-containing complex NODE (for Nanog and Oct4 associated deacetylase). Interestingly, NODE contained histone deacetylase (HDAC) activity that seemed to be comparable to NuRD, and retained its association with Nanog and Oct4 in Mbd3−/− ES cells. In contrast to Mbd3 loss-of-function, knockdown of NODE subunits led to increased expression of developmentally regulated genes and ES-cell differentiation. Our data collectively suggest that Nanog and Oct4 associate with unique repressor complexes on their target genes to control ES cell fate.

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Figure 1: Identification of the endogenous Nanog protein complexes from mouse ES cells.
Figure 2: Nanog and Oct4 recruit multiple repressor complexes.
Figure 3: Nanog and Oct4 repression complexes contain HDAC activity but a low stoichiometric amount of Mbd3.
Figure 4: Co-occupancy of Nanog, Oct4 and Hdac2 on Nanog target genes is independent of Mbd3.
Figure 5: Effects of Mta1, Mta2 or Mbd3 RNAi knockdown on expression of Nanog and Oct4 target genes and ES cell self-renewal and differentiation.

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Acknowledgements

We thank Amin Safari, Xueping Xu and Ok-Hee Lee for technical assistance. We also thank Yaoyun Liang and Xinhua Feng for helping us with quantitative PCR analysis. We thank Brian Hendrich for providing the Mbd3−/− ES cells. This work was supported by NIH grants GM69572 and GM81627 to Z.S and DK73524 to A.J.C. D.L. is supported by American Heart Association. Z.S. is a Leukemia and Lymphoma Society scholar.

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Author notes

  1. Peili Gu

    Present address: Current address: Department of Cancer Genetics, University of Texas, M.D. Anderson Cancer Center, Houston, TX 77030, USA.,

  2. Jiemin Wong

    Present address: Current address: The Institute of Biomedical Sciences, College of Life Sciences, East China Normal University, Shanghai, China.,

  3. Jiancong Liang and Ma Wan: These authors contributed equally to this work.

Authors and Affiliations

  1. Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, 77030, Texas, USA

    Jiancong Liang, Ma Wan, Yi Zhang, Huawei Xin, Sung Yun Jung, Jun Qin, Dan Liu & Zhou Songyang

  2. Department of Molecular and Cellular Biology, Baylor College of Medicine, One Baylor Plaza, Houston, 77030, Texas, USA

    Peili Gu, Jun Qin, Jiemin Wong & Austin J. Cooney

  3. Stem Cells and Regenerative Medicine Center, Baylor College of Medicine, One Baylor Plaza, Houston, 77030, Texas, USA

    Austin J. Cooney & Zhou Songyang

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Contributions

J.L., M.W., Y.Z., P.G., H.X., S.Y.J., J.Q. and D.L. performed the experiments; J.W. and A.J.C. provided reagents; Z.S. provided advice on the experimental design and wrote the manuscript. All authors commented on the manuscript.

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Correspondence to Zhou Songyang.

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Supplementary Figures S1, S2, S3, S4, S5, S6 and Supplementary Tables S1, S2 (PDF 748 kb)

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Liang, J., Wan, M., Zhang, Y. et al. Nanog and Oct4 associate with unique transcriptional repression complexes in embryonic stem cells. Nat Cell Biol 10, 731–739 (2008). https://doi.org/10.1038/ncb1736

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