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Chk2 activates E2F-1 in response to DNA damage

Nature Cell Biology volume 5, pages 401–409 (2003)Cite this article

Abstract

The E2F-1 transcription factor is regulated during cell cycle progression and induced by cellular stress, such as DNA damage. We report that checkpoint kinase 2 (Chk2) regulates E2F-1 activity in response to the DNA-damaging agent etoposide. A Chk2 consensus phosphorylation site in E2F-1 is phosphorylated in response to DNA damage, resulting in protein stabilization, increased half-life, transcriptional activation and localization of phosphorylated E2F-1 to discrete nuclear structures. Expression of a dominant-negative Chk2 mutant blocks induction of E2F-1 and prevents E2F-1-dependent apoptosis. Moreover, E2F-1 is resistant to induction by etoposide in tumour cells expressing mutant chk2. Therefore, Chk2 phosphorylates and activates E2F-1 in response to DNA damage, resulting in apoptosis. These results suggest a role for E2F-1 in checkpoint control and provide a plausible explanation for the tumour suppressor activity of E2F-1.

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Figure 1: E2F-1 is stabilised after DNA damage and requires Chk2 kinase.
Figure 2: Chk2 phosphorylates E2F-1 on Ser 364 in vitro.
Figure 3: E2F-1 is phosphorylated on Ser 364 in vivo.
Figure 4: Dependence on Ser 364 for E2F-1 stabilisation in response to DNA damage.
Figure 5: Role of Chk2 in the stabilisation of E2F-1 after DNA damage.
Figure 6: Localization of E2F-1-Ser 364-P in normal cells.
Figure 7: Detection of E2F-1-Ser 364-P in etoposide-treated cells.
Figure 8: Apoptosis and cell cycle arrest through E2F-1 requires Chk2.

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Acknowledgements

We thank J. Zhu and T. Halazonetis for materials. This work was supported by the Leukaemia Research Fund (C.S.) and the Medical Research Council.

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  1. Division of Biochemistry and Molecular Biology, Davidson Building, University of Glasgow, Glasgow, G12 8QQ, UK

    Craig Stevens, Linda Smith & Nicholas B. La Thangue

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Correspondence to Nicholas B. La Thangue.

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Supplementary Figures

Figure S1 Purification and specificity of phospho-specific anti-P-S364. (PDF 48 kb)

Figure S2 Recombinant E2F-1 or S364A (1µg) was incubated with recombinant Chk2 as indicated, in the presence of either the phosphorylated (track 5) or nonphosphorylated (track 6) S364 peptide (about 2µg).

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Stevens, C., Smith, L. & La Thangue, N. Chk2 activates E2F-1 in response to DNA damage. Nat Cell Biol 5, 401–409 (2003). https://doi.org/10.1038/ncb974

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