Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Triggering neural differentiation of ES cells by subtype switching of importin-α

A Corrigendum to this article was published on 01 April 2007

Abstract

Nuclear proteins are selectively imported into the nucleus by transport factors such as importin-α and importin-β1,2. Here, we show that the expression of importin-α subtypes is strictly regulated during neural differentiation of mouse embryonic stem (ES) cells, and that the switching of importin-α subtype expression is critical for neural differentiation. Moreover, reproducing the switching of importin-α subtype expression in undifferentiated ES cells induced neural differentiation in the presence of leukaemia inhibitory factor (LIF) and serum, coordinated with the regulated expression of Oct3/4, Brn2 and SOX2, which are involved in ES–neural identity determination. These transcription factors were selectively imported into the nucleus by specific subtypes of importin-α. Thus, importin-α subtype switching has a major impact on cell differentiation through the regulated nuclear import of a specific set of transcription factors. This is the first study to propose that transport factors should be considered as major players in cell-fate determination.

This is a preview of subscription content, access via your institution

Access options

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

Figure 1: Expression profiles of importin-α subtypes during the retinoic acid-induced neural differentiation of ES cells.
Figure 2: The effects of overexpression of importin-α1 and suppression of importin-α5 expression on neural differentiation of ES cells.
Figure 3: The effects of downregulation of importin-α1 expression and upregulation of importin-α5 expression on ES cells cultured in LIF+ medium.
Figure 4: Differential nuclear import of Oct3/4, Brn2, SOX2 and Oct6 by importin-α1, -α3 and -α5.
Figure 5: A schematic representation of a model for cell-fate determination by the interdependent regulation of nuclear transport machineries and transcription factors.

Similar content being viewed by others

References

  1. Görlich, D. & Mattaj, I. W. Nucleocytoplasmic transport. Science 271, 1513–1518 (1996).

    Article  Google Scholar 

  2. Goldfarb, D. S. et al. Importin-α: a multipurpose nuclear-transport receptor. Trends Cell Biol. 14, 505–514 (2004).

    Article  CAS  Google Scholar 

  3. Yoneda, Y. Nucleocytoplasmic protein traffic and its significance to cell function. Genes Cells 5, 777–787 (2000).

    Article  CAS  Google Scholar 

  4. Köhler, M. et al. Evidence for distinct substrate specificities of importin-α family members in nuclear protein import. Mol. Cell. Biol. 11, 7782–7791 (1999).

    Article  Google Scholar 

  5. Kawasaki, H. et al. Induction of midbrain dopaminergic neurons from ES cells by stromal cell-derived inducing activity. Neuron 28, 31–40 (2000).

    Article  CAS  Google Scholar 

  6. Ying, Q. L. et al. Conversion of embryonic stem cells into neuroectodermal precursors in adherent monoculture. Nature Biotechnol. 2, 183–186 (2003).

    Article  Google Scholar 

  7. Tanaka, S. et al. Interplay of SOX and POU factors in regulation of the Nestin gene in neural primordial cells. Mol. Cell Biol. 24, 8834–8846 (2004).

    Article  CAS  Google Scholar 

  8. Kamachi, Y., Uchikawa, M. & Kondoh, H. Pairing SOX off: with partners in the regulation of embryonic development. Trends Genet. 16, 182–187 (2000).

    Article  CAS  Google Scholar 

  9. Yuan, H., Corbi, N., Basilico, C. & Dailey, L. Developmental-specific activity of the FGF-4 enhancer requires the synergistic action of Sox2 and Oct-3. Genes Dev. 9, 2635–2645 (1995).

    Article  CAS  Google Scholar 

  10. Chew, J. L. et. al. Reciprocal transcriptional regulation of Pou5f1 and Sox2 via the Oct4/Sox2 complex in embryonic stem cells. Mol Cell Biol. 25, 6031–6046 (2005).

    Article  CAS  Google Scholar 

  11. Rodda, D. J. et. al. Transcriptional regulation of nanog by OCT4 and SOX2. J. Biol. Chem. 280, 24731–24737 (2005).

    CAS  Google Scholar 

  12. Kuroda, T. et. al. Octamer and Sox elements are required for transcriptional cis regulation of Nanog gene expression. Mol. Cell Biol. 25, 2475–2485 (2005).

    Article  CAS  Google Scholar 

  13. Pan, G. et al. Identification of a nuclear localization signal in OCT4 and generation of a dominant negative mutant by its ablation. J. Biol. Chem. 279, 37013–37020 (2004).

    Article  CAS  Google Scholar 

  14. Sock, E., Enderich, J., Rosenfeld, M. G. & Wegner, M. Identification of the nuclear localization signal of the POU domain protein Tst-1/Oct6. J. Biol. Chem. 271, 17512–17528 (1996).

    Article  CAS  Google Scholar 

  15. Argentaro, A. et al. A SOX9 defect of calmodulin-dependent nuclear import in campomelic dysplasia/autosomal sex reversal. J. Biol. Chem. 278, 33839–33847 (2003).

    Article  CAS  Google Scholar 

  16. Baranek, C., Sock, E. & Wegner, M. The POU protein Oct-6 is a nucleocytoplasmic shuttling protein. Nucleic Acids Res. 33, 6277–6286 (2005).

    Article  CAS  Google Scholar 

  17. Ilia, M., Bazigou, E. & Price, J. Expression of the POU domain transcription factor, Oct-6, is attenuated in the adult mouse telencephalon, but increased by neurotoxic damage. Exp. Neurol. 181, 159–169 (2003).

    Article  CAS  Google Scholar 

  18. Okumura-Nakanishi, S., Saito, M., Niwa, H. & Ishikawa, F. Oct-3/4 and Sox2 regulate Oct-3/4 gene in embryonic stem cells. J. Biol. Chem. 280, 5307–5317 (2005).

    Article  CAS  Google Scholar 

  19. Tomioka, M. et al. Identification of Sox-2 regulatory region which is under the control of Oct-3/4-Sox-2 complex. Nucleic Acids Res. 30, 3202–3213 (2002).

    Article  CAS  Google Scholar 

  20. Boyer, L. A. et al. Core transcriptional regulatory circuitry in human embryonic stem cells. Cell. 122, 947–956 (2005).

    Article  CAS  Google Scholar 

  21. Niwa, H., Miyazaki, J. & Smith, A. G. Quantitative expression of Oct3/4 defines differentiation or self-renewal of ES cells. Nature Genet. 24, 372–376 (2000).

    Article  CAS  Google Scholar 

  22. Rudnicki, M. A. & McBurney, M. W. in Teratocarcinomas and Embryonic Stem Cells: A partial Approach. (ed. Robertson E. J.) 19–49 (IRL Press, Oxford, 1987).

    Google Scholar 

  23. Kamikubo, Y. et al. Specific monoclonal antibody against nuclear import factor, importin-α1/Rch1. Hybrid Hybridomics 5, 301–304 (2004).

    Article  Google Scholar 

  24. Sakaguchi, T. et al. Generation of a rat monoclonal antibody specific for importin-α 3/Qip1. Hybrid Hybridomics. 6, 397–400 (2003).

    Article  Google Scholar 

  25. Imamoto, N. et al. Antibodies against 70-kD heat shock cognate protein inhibit mediated nuclear import of karyophilic proteins J. Cell Biol. 119, 1047–1061 (1992).

    Article  CAS  Google Scholar 

  26. Yoneda, Y., Imamoto-Sonobe, N., Yamaizumi, M. & Uchida, T. Reversible inhibition of protein import into the nucleus by wheat germ agglutinin injected into cultured cells. Exp. Cell Res. 173, 586–595 (1987).

    Article  CAS  Google Scholar 

  27. Adam, S. A., Marr, R. S. & Gerace, L. Nuclear protein import in permeabilized mammalian cells requires soluble cytoplasmic factors. J. Cell Biol. 111, 807–816 (1990).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank H. Niwa and T. Tachibana for providing the ES cell line and antibodies, A. G. Smith for the suggestion of using spreading culture conditions, and M. Raff, S. Nagata and T. Nakano for helpful comments. We also thank Y. Yoshida and T. Yamashita for help during the experiments. This work was supported by grants from the Ministry of Education, Culture, Sports, Science, and Technology of Japan and the Human Frontier Science Program.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Yoshihiro Yoneda.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary figures S1, S2, S3, S4 and Supplementary Table S1 (PDF 749 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yasuhara, N., Shibazaki, N., Tanaka, S. et al. Triggering neural differentiation of ES cells by subtype switching of importin-α. Nat Cell Biol 9, 72–79 (2007). https://doi.org/10.1038/ncb1521

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ncb1521

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing