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Radiation-mediated proteolysis of CDT1 by CUL4–ROC1 and CSN complexes constitutes a new checkpoint

Nature Cell Biology volume 5, pages 1008–1015 (2003)Cite this article

An Erratum to this article was published on 01 December 2003

Abstract

Genomic integrity is maintained by checkpoints that guard against undesired replication after DNA damage. Here, we show that CDT1, a licensing factor of the pre-replication complex (preRC)1,2,3, is rapidly proteolysed after UV- or γ-irradiation. The preRC assembles on replication origins at the end of mitosis and during G1 to license DNA for replication in S phase. Once the origin recognition complex (ORC) binds to origins, CDC6 and CDT1 associate with ORC and promote loading of the MCM2-7 proteins onto chromatin, generating the preRC. We show that radiation-mediated CDT1 proteolysis is independent of ATM and CHK2 and can occur in G1-phase cells. Loss of the COP9–signalosome (CSN) or CUL4–ROC1 complexes completely suppresses CDT1 proteolysis. CDT1 is specifically polyubiquitinated by CUL4 complexes and the interaction between CDT1 and CUL4 is regulated in part by γ-irradiation. Our study reveals an evolutionarily conserved and uncharacterized G1 checkpoint that induces CDT1 proteolysis by the CUL4–ROC1 ubiquitin E3 ligase and CSN complexes in response to DNA damage.

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Figure 1: CDT1 is rapidly proteolysed after UV- or γ-irradiation.
Figure 2: CDT1 proteolysis after γ-irradiation is independent of ATM, ATR, CHK1 and CHK2 kinases.
Figure 3: CDT1 proteolysis after irradiation occurs in G1 independent of DNA replication.
Figure 4: Loss of CSN, ROC1a, or CUL4 blocks CDT1 degradation after irradiation.
Figure 5: CDT1 is a direct ubiquitination substrate of CUL4 E3 ligases.

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Acknowledgements

This work was supported by grants from the National Institutes of Health (CA72878) and the Department of the US Army (DAMD17-00-1-0511) to H. Z. L. A. H. was supported by a Predoctoral Training Program in Breast Cancer Research (DAMD17-99-1-9461) from the US Army Medical Research and Materiel Command.

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  1. Department of Genetics, Yale University School of Medicine, 333 Cedar Street, New Haven, 06520, CT, USA

    Leigh Ann A. Higa, Ivailo S. Mihaylov, Damon P. Banks, Jianyu Zheng & Hui Zhang

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Higa, L., Mihaylov, I., Banks, D. et al. Radiation-mediated proteolysis of CDT1 by CUL4–ROC1 and CSN complexes constitutes a new checkpoint. Nat Cell Biol 5, 1008–1015 (2003). https://doi.org/10.1038/ncb1061

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