Published online
doi:10.1083/jcb.200708210
The Journal of Cell Biology, Vol. 181, No. 2, 213-226
The Rockefeller University Press, 0021-9525 $30.00
© Melander et al.
Phosphorylation of SDT repeats in the MDC1 N terminus triggers retention of NBS1 at the DNA damage–modified chromatin
Fredrik Melander1,2,
Simon Bekker-Jensen1,2,
Jacob Falck1,2,
Jiri Bartek1,2,
Niels Mailand1,2, and
Jiri Lukas1,2
1 Institute of Cancer Biology and 2 Centre for Genotoxic Stress Research, Danish Cancer Society, DK-2100 Copenhagen, Denmark
Correspondence to Jiri Bartek: jb{at}cancer.dk; or Jiri Lukas: jil{at}cancer.dk
DNA double-strand breaks (DSBs) trigger accumulation of the MRE11–RAD50–Nijmegen breakage syndrome 1 (NBS1 [MRN]) complex, whose retention on the DSB-flanking chromatin facilitates survival. Chromatin retention of MRN requires the MDC1 adaptor protein, but the mechanism behind the MRN–MDC1 interaction is unknown. We show that the NBS1 subunit of MRN interacts with the MDC1 N terminus enriched in Ser-Asp-Thr (SDT) repeats. This interaction was constitutive and mediated by binding between the phosphorylated SDT repeats of MDC1 and the phosphate-binding forkhead-associated domain of NBS1. Phosphorylation of the SDT repeats by casein kinase 2 (CK2) was sufficient to trigger MDC1–NBS1 interaction in vitro, and MDC1 associated with CK2 activity in cells. Inhibition of CK2 reduced SDT phosphorylation in vivo, and disruption of the SDT-associated phosphoacceptor sites prevented the retention of NBS1 at DSBs. Together, these data suggest that phosphorylation of the SDT repeats in the MDC1 N terminus functions to recruit NBS1 and, thereby, increases the local concentration of MRN at the sites of chromosomal breakage.
F. Melander and S. Bekker-Jensen contributed equally to this paper.
J. Falck's present address is Novo Nordisk A/S, DK-2880 Bagsverd, Denmark.
Abbreviations used in this paper: ATM, ataxia telangiectasia mutated; BRCT, BRCA1 C terminal; CK2, casein kinase 2; DSB, double-strand break; FHA, forkhead-associated; IR, ionizing radiation; MRN, MRE11–RAD50–NBS1; NBS, Nijmegen breakage syndrome; SDT, Ser-Asp-Thr; shRNA, short hairpin RNA; WT, wild type.
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