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Regulation of the activation of the Fanconi anemia pathway by the p21 cyclin-dependent kinase inhibitor

Oncogene volume 31, pages 366–375 (2012)Cite this article

Abstract

Fanconi anemia (FA) is a rare disease characterized by congenital defects, progressive bone marrow failure and heightened cancer susceptibility. The FA proteins, BRCA1 and FANCD1/BRCA2 function cooperatively in the FA-BRCA pathway to repair damaged DNA. Activation of the FA-BRCA pathway occurs via the monoubiquitination of the FANCD2 and FANCI proteins, targeting these proteins to discrete nuclear foci where they function in DNA repair. The cellular regulation of FANCD2/I monoubiquitination, however, remains poorly understood. In this study, we have examined the roles of the p53 tumor suppressor protein, as well as its downstream target, the p21Cip1/Waf1 cyclin-dependent kinase inhibitor, in the regulation of the activation of the FA-BRCA pathway. We demonstrate that, in contrast to p53, p21 has a major role in the regulation of the activation of the FA-BRCA pathway: p21 promotes S-phase and DNA damage-inducible FANCD2/I monoubiquitination and nuclear foci formation. Several lines of evidence establish that this effect is not a consequence of a defective G1–S checkpoint or altered cell-cycle progression in the absence of p21. Instead, we demonstrate that p21 is required for the transcriptional repression of the USP1 deubiquitinating enzyme upon exposure to DNA-damaging agents. In the absence of p21, persistent USP1 expression precludes the DNA damage-inducible accumulation of monoubiquitinated FANCD2 and FANCI. Consequently, p21−/− cells exhibit increased levels of mitomycin C-inducible complex chromosomal aberrations and elevated γH2AX nuclear foci formation. Our results demonstrate that p21 has a critical role in the regulation of the activation of the FA-BRCA pathway and suggest a broader role for p21 in the orchestration of DNA repair processes following exposure to DNA crosslinking agents.

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Acknowledgements

We thank the members of the Howlett laboratory, Paul R Andreassen, Matthew Stoner and Patrick Sung for helpful discussions. We thank Tony T Huang and Patrick Sung for the anti-USP1 and anti-FANCI antibodies, respectively. We thank Bert Vogelstein for cells. This work was supported by a Leukemia Research Foundation New Investigator grant (NGH), RI-INBRE Grant P20RR016457-09 from the National Center for Research Resources (NGH) and National Institutes of Health/National Heart, Lung and Blood Institute Grant R21HL095991-01 (NGH).

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  1. Department of Cell and Molecular Biology, University of Rhode Island, Kingston, RI, USA

    M A Rego, J A Harney, M Mauro, M Shen & N G Howlett

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  1. M A Rego
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Correspondence to N G Howlett.

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Rego, M., Harney, J., Mauro, M. et al. Regulation of the activation of the Fanconi anemia pathway by the p21 cyclin-dependent kinase inhibitor. Oncogene 31, 366–375 (2012). https://doi.org/10.1038/onc.2011.237

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