Functional interaction between FOXO3a and ATM regulates DNA damage response

  • A Corrigendum to this article was published on 01 November 2009

Abstract

The maintenance of genomic stability in cells is relentlessly challenged by environmental stresses that induce DNA breaks, which activate the DNA-damage pathway mediated by ataxia-telangiectasia mutated (ATM) and its downstream mediators to control damage-induced cell-cycle checkpoints and DNA repair1,2,3. Here, we show that FOXO3a interacts with ATM to promote phosphorylation of ATM at Ser 1981 and prompting its downstream mediators to form nuclear foci in response to DNA damage. Silencing FOXO3a in cells abrogates the formation of ATM-pS1981 and phospho-histone H2AX foci after DNA damage. Increasing FOXO3a in cells promotes ATM-regulated signalling, the intra-S-phase or G2–M cell-cycle checkpoints, and the repair of damaged DNA, whereas cells lacking FOXO3a did not trigger the DNA-repair mechanism after DNA damage. The carboxy-terminal domain of FOXO3a binds to the FAT domain of ATM, thereby contributing to the activation of ATM. These results suggest that ATM may be regulated directly by FOXO3a in the DNA-damage response.

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Figure 1: Silencing FOXO3a results in defective intra-S-phase and G2–M phase cell-cycle checkpoints.
Figure 2: FOXO3a may be colocalized with γ-H2AX and ATM-pS1981 to form nuclear foci in cells treated with ionizing radiation or CPT.
Figure 3: FOXO3a is colocalized with ATM-pSer1981 or γ-H2AX in primary tumour tissues in vivo, associated with these phospho-proteins in the nucleus in cells treated with CPT, and involved in ATM autophosphorylation of Ser 1981.
Figure 4: FOXO3a promotes the repair of damaged DNA.
Figure 5: The carboxy-terminal domain of FOXO3a binds to the FAT domain of ATM in vitro and in vivo.

Change history

  • 01 October 2009

    In the version of this article initially published, the top left panel in Fig. 4c was incorrect. This error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We thank M. B. Kastan, R. Depinho, P. Coffer, S. Miyamoto, A. J. Fornace Jr. and K. K. Khanna for generously providing reagents. This work was supported in part by R01 grant CA113859 (M.C.-T.H.) from the National Cancer Institute (NCI), National Institutes of Health; grants BCTR0504415 from the Susan G. Komen Breast Cancer Foundation, BC045295 from the U.S. Department of Defense Breast Cancer Research Program, and a grant from the Texas Advanced Research Program (M.C.-T.H.); and Cancer Center Support Grant CA16772 from the NCI. The sponsors had no role in the design, conduct or reporting of the study.

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W.B.T. performed the majority of experiments, including the cell-cycle checkpoint, siRNA knockdown, coimmunoprecipitation, immunoblotting, immunofluorescence confocal microscopy, tumour staining, GST pulldown, immunocomplex kinase assays and data analysis. Y.M.C. performed all the analysis of DNA damage by the comet assays and quantification of immunofluorescence images. Y.T. conducted quantitative real-time PCR analyses, mammalian and yeast two-hybrid assays. Z.X. provided crucial assistance in generating DNA constructs, cell lines, GST fusion proteins and performing luciferase and PCR analyses. M.C.-T.H. generated DNA constructs and cell lines, designed and coordinated all experimental approaches, drafted and revised the entire manuscript.

Corresponding author

Correspondence to Mickey C.-T. Hu.

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Supplementary Figures S1, S2, S3, S4, S5, S6 and Supplementary Text (PDF 2664 kb)

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Tsai, W., Chung, Y., Takahashi, Y. et al. Functional interaction between FOXO3a and ATM regulates DNA damage response. Nat Cell Biol 10, 460–467 (2008). https://doi.org/10.1038/ncb1709

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