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.

  • Letter
  • Published:

Degradation of Id2 by the anaphase-promoting complex couples cell cycle exit and axonal growth

Abstract

In the developing nervous system, Id2 (inhibitor of DNA binding 2, also known as inhibitor of differentiation 2) enhances cell proliferation, promotes tumour progression and inhibits the activity of neurogenic basic helix–loop–helix (bHLH) transcription factors1,2. The anaphase promoting complex/cyclosome and its activator Cdh1 (APC/CCdh1) restrains axonal growth but the targets of APC/CCdh1 in neurons are unknown3,4,5. Id2 and other members of the Id family are very unstable proteins that are eliminated as cells enter the quiescent state, but how they are targeted for degradation has remained elusive6,7. Here we show that Id2 interacts with the core subunits of APC/C and Cdh1 in primary neurons. APC/CCdh1 targets Id2 for degradation through a destruction box motif (D box) that is conserved in Id1 and Id4. Depletion of Cdh1 stabilizes Id proteins in neurons, whereas Id2 D-box mutants are impaired for Cdh1 binding and remain stable in cells that exit from the cell cycle and contain active APC/CCdh1. Mutants of the Id2 D box enhance axonal growth in cerebellar granule neurons in vitro and in the context of the cerebellar cortex, and overcome the myelin inhibitory signals for growth. Conversely, activation of bHLH transcription factors induces a cluster of genes with potent axonal inhibitory functions including the gene coding for the Nogo receptor, a key transducer of myelin inhibition. Degradation of Id2 in neurons permits the accumulation of the Nogo receptor, thereby linking APC/CCdh1 activity with bHLH target genes for the inhibition of axonal growth. These findings indicate that deregulated Id activity might be useful to reprogramme quiescent neurons into the axonal growth mode.

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: Id2 is a substrate of APC/CCdh1.
Figure 2: D-box-dependent degradation of Id proteins by APC/CCdh1.
Figure 3: An APC/C Cdh1 –Id–bHLH pathway controls axonal growth.
Figure 4: Significance of APC/C Cdh1 -mediated degradation of Id2 for axonal growth.

Similar content being viewed by others

References

  1. Iavarone, A. & Lasorella, A. Id proteins in neural cancer. Cancer Lett. 204, 189–196 (2004)

    Article  CAS  PubMed  Google Scholar 

  2. Perk, J., Iavarone, A. & Benezra, R. Id family of helix–loop–helix proteins in cancer. Nature Rev. Cancer 5, 603–614 (2005)

    Article  CAS  Google Scholar 

  3. Konishi, Y., Stegmuller, J., Matsuda, T., Bonni, S. & Bonni, A. Cdh1-APC controls axonal growth and patterning in the mammalian brain. Science 303, 1026–1030 (2004)

    Article  ADS  CAS  PubMed  Google Scholar 

  4. Peters, J. M. The anaphase-promoting complex: proteolysis in mitosis and beyond. Mol. Cell 9, 931–943 (2002)

    Article  CAS  PubMed  Google Scholar 

  5. Stegmuller, J. & Bonni, A. Moving past proliferation: new roles for Cdh1-APC in postmitotic neurons. Trends Neurosci. 28, 596–601 (2005)

    Article  PubMed  Google Scholar 

  6. Bounpheng, M. A., Dimas, J. J., Dodds, S. G. & Christy, B. A. Degradation of Id proteins by the ubiquitin–proteasome pathway. FASEB J. 13, 2257–2264 (1999)

    Article  CAS  PubMed  Google Scholar 

  7. Fajerman, I., Schwartz, A. L. & Ciechanover, A. Degradation of the Id2 developmental regulator: targeting via N-terminal ubiquitination. Biochem. Biophys. Res. Commun. 314, 505–512 (2004)

    Article  CAS  PubMed  Google Scholar 

  8. Wainwright, L. J., Lasorella, A. & Iavarone, A. Distinct mechanisms of cell cycle arrest control the decision between differentiation and senescence in human neuroblastoma cells. Proc. Natl Acad. Sci. USA 98, 9396–9400 (2001)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  9. Reimann, J. D. et al. Emi1 is a mitotic regulator that interacts with Cdc20 and inhibits the anaphase promoting complex. Cell 105, 645–655 (2001)

    Article  CAS  PubMed  Google Scholar 

  10. Harper, J. W., Burton, J. L. & Solomon, M. J. The anaphase-promoting complex: it's not just for mitosis any more. Genes Dev. 16, 2179–2206 (2002)

    Article  CAS  PubMed  Google Scholar 

  11. Gieffers, C., Peters, B. H., Kramer, E. R., Dotti, C. G. & Peters, J. M. Expression of the CDH1-associated form of the anaphase-promoting complex in postmitotic neurons. Proc. Natl Acad. Sci. USA 96, 11317–11322 (1999)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  12. Powell, S. K., Rivas, R. J., Rodriguez-Boulan, E. & Hatten, M. E. Development of polarity in cerebellar granule neurons. J. Neurobiol. 32, 223–236 (1997)

    Article  CAS  PubMed  Google Scholar 

  13. Barallobre, M. J., Pascual, M., Del Rio, J. A. & Soriano, E. The Netrin family of guidance factors: emphasis on Netrin-1 signalling. Brain Res. Brain Res. Rev. 49, 22–47 (2005)

    Article  CAS  PubMed  Google Scholar 

  14. Fiore, R. & Puschel, A. W. The function of semaphorins during nervous system development. Front. Biosci. 8, s484–s499 (2003)

    Article  CAS  PubMed  Google Scholar 

  15. Sestan, N., Artavanis-Tsakonas, S. & Rakic, P. Contact-dependent inhibition of cortical neurite growth mediated by notch signaling. Science 286, 741–746 (1999)

    Article  CAS  PubMed  Google Scholar 

  16. Schwab, M. E. Nogo and axon regeneration. Curr. Opin. Neurobiol. 14, 118–124 (2004)

    Article  CAS  PubMed  Google Scholar 

  17. Spencer, T., Domeniconi, M., Cao, Z. & Filbin, M. T. New roles for old proteins in adult CNS axonal regeneration. Curr. Opin. Neurobiol. 13, 133–139 (2003)

    Article  CAS  PubMed  Google Scholar 

  18. Lesuisse, C. & Martin, L. J. Long-term culture of mouse cortical neurons as a model for neuronal development, aging, and death. J. Neurobiol. 51, 9–23 (2002)

    Article  PubMed  Google Scholar 

  19. Zhou, Y., Ching, Y. P., Chun, A. C. & Jin, D. Y. Nuclear localization of the cell cycle regulator CDH1 and its regulation by phosphorylation. J. Biol. Chem. 278, 12530–12536 (2003)

    Article  CAS  PubMed  Google Scholar 

  20. Wirth, K. G. et al. Loss of the anaphase-promoting complex in quiescent cells causes unscheduled hepatocyte proliferation. Genes Dev. 18, 88–98 (2004)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Baghdoyan, S. et al. Id2 reverses cell cycle arrest induced by γ-irradiation in human HaCaT keratinocytes. J. Biol. Chem. 280, 15836–15841 (2005)

    Article  CAS  PubMed  Google Scholar 

  22. Kowanetz, M., Valcourt, U., Bergstrom, R., Heldin, C. H. & Moustakas, A. Id2 and Id3 define the potency of cell proliferation and differentiation responses to transforming growth factor β and bone morphogenetic protein. Mol. Cell. Biol. 24, 4241–4254 (2004)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Lasorella, A. et al. Id2 is critical for cellular proliferation and is the oncogenic effector of N-myc in human neuroblastoma. Cancer Res. 62, 301–306 (2002)

    CAS  PubMed  Google Scholar 

  24. Chaudhary, J., Sadler-Riggleman, I., Ague, J. M. & Skinner, M. K. The helix–loop–helix inhibitor of differentiation (ID) proteins induce post-mitotic terminally differentiated sertoli cells to re-enter the cell cycle and proliferate. Biol. Reprod. 72, 1205–1217 (2005)

    Article  CAS  PubMed  Google Scholar 

  25. Encinas, M. et al. Sequential treatment of SH-SY5Y cells with retinoic acid and brain-derived neurotrophic factor gives rise to fully differentiated, neurotrophic factor-dependent, human neuron-like cells. J. Neurochem. 75, 991–1003 (2000)

    Article  CAS  PubMed  Google Scholar 

  26. Aarts, M. et al. A key role for TRPM7 channels in anoxic neuronal death. Cell 115, 863–877 (2003)

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank S. Gygi and N. Sherman for the identification of Id2-associated proteins by mass spectrometry; P. K. Jackson for providing us with purified MBP-Emi1; and W. G. Kaelin and J. Lukas for the U2OS cells conditionally expressing Myc-Cdh1. This work was supported by grants from the National Institutes of Health to A.L., A.I., A.B. and M.P., from the Charlotte Geyer Foundation to A.I., and from the Christopher Reeve Paralysis Foundation to A.B. J.S. and D.G. are supported by Deutsche Forschungsgemeinschaft and Emerald Foundation grants, respectively.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Antonio Iavarone.

Ethics declarations

Competing interests

The microarray data have been deposited in the ArrayExpress database (http://www.ebi.ac.uk/arrayexpress/query/entry) under accession number E-MEXP-413. Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.

Supplementary information

Supplementary Methods

This file contains additional details of the methods used in this study. (PDF 117 kb)

Supplementary Figure Legends

This file contains text to accompany the below Supplementary Figures. (DOC 57 kb)

Supplementary Figure 1

Id2 is degraded by APC/CCdh1 and binds to the core subunits of APC/C and Cdh1. (PDF 97 kb)

Supplementary Figure 2

Id2 is unstable in cells withdrawing the cell cycle. (PDF 31 kb)

Supplementary Figure 3

Id2 protein stability in quiescent cells is regulated by Cdh1. (PDF 90 kb)

Supplementary Figure 4

Degradation of Id protein by APC/CCdh1 is dependent on D box. (PDF 42 kb)

Supplementary Figure 5

Dual interaction modules of Id2 with Cdh1 and core APC/C. (PDF 66 kb)

Supplementary Figure 6

Id2 does not affect the integrity and activity of APC/CCdh1 (PDF 56 kb)

Supplementary Figure 7

Id2 is a target of APC/CCdh1 for axonal growth. (PDF 414 kb)

Supplementary Figure 8

Degradation resistant Id2 promotes axonal growth of cerebellar granule neurons in vitro and in vivo. (PDF 234 kb)

Supplementary Figure 9

A functional Cdh1-Id-bHLH pathway controls axonal growth. (PDF 245 kb)

Supplementary Figure 10

Silencing of Cdh1 in cortical neurons stabilizes Id2 and Id1. (PDF 21 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lasorella, A., Stegmüller, J., Guardavaccaro, D. et al. Degradation of Id2 by the anaphase-promoting complex couples cell cycle exit and axonal growth. Nature 442, 471–474 (2006). https://doi.org/10.1038/nature04895

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature04895

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

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