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.

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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.

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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.

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Correspondence to Antonio Iavarone.

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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)

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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

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