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MDC1 is coupled to activated CHK2 in mammalian DNA damage response pathways

Nature volume 421, pages 957–961 (2003)Cite this article

Abstract

Forkhead-homology-associated (FHA) domains function as protein–protein modules that recognize phosphorylated serine/threonine motifs1,2,3,4,5. Interactions between FHA domains and phosphorylated proteins are thought to have essential roles in the transduction of DNA damage signals; however, it is unclear how FHA-domain-containing proteins participate in mammalian DNA damage responses. Here we report that a FHA-domain-containing protein—mediator of DNA damage checkpoint protein 1 (MDC1; previously known as KIAA0170)—is involved in DNA damage responses. MDC1 localizes to sites of DNA breaks and associates with CHK2 after DNA damage. This association is mediated by the MDC1 FHA domain and the phosphorylated Thr 68 of CHK2. Furthermore, MDC1 is phosphorylated in an ATM/CHK2-dependent manner after DNA damage, suggesting that MDC1 may function in the ATM–CHK2 pathway. Consistent with this hypothesis, suppression of MDC1 expression results in defective S-phase checkpoint and reduced apoptosis in response to DNA damage, which can be restored by the expression of wild-type MDC1 but not MDC1 with a deleted FHA domain. Suppression of MDC1 expression results in decreased p53 stabilization in response to DNA damage. These results suggest that MDC1 is recruited through its FHA domain to the activated CHK2, and has a critical role in CHK2-mediated DNA damage responses.

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Figure 1: MDC1is phosphorylated and relocalizes to nuclear foci after DNA damage.
Figure 2: MDC1 interacts with CHK2 through the MDC1 FHA domain and the phosphorylated Thr 68 of CHK2.
Figure 3: MDC1 is required for CHK2-mediated DNA damage responses.
Figure 4: The functional interaction of CHK2 and MDC1 is required for CHK2-mediated DNA damage responses.

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Acknowledgements

We thank L. Wang for technical support. We also thank Mayo Protein Core facility for synthesis of peptides. We are grateful to L. Karnitz and S. Kaufmann and members of the Chen and Karnitz laboratories for discussions and ongoing technical support. This work is supported in part by grants from the National Institute of Health, the Prospect Creek Foundation and the Breast Cancer Research Foundation. J.C. is a recipient of a DOD breast cancer career development award.

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  1. Department of Oncology, Mayo Foundation, Rochester, Minnesota, 55905, USA

    Zhenkun Lou, Katherine Minter-Dykhouse, Xianglin Wu & Junjie Chen

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Correspondence to Junjie Chen.

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Lou, Z., Minter-Dykhouse, K., Wu, X. et al. MDC1 is coupled to activated CHK2 in mammalian DNA damage response pathways. Nature 421, 957–961 (2003). https://doi.org/10.1038/nature01447

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