Phospho-dependent Interactions Between NBS1 and MDC1 Mediate Chromatin Retention of the MRN Complex at Sites of DNA Damage

Overview

Journal EMBO Rep

Specialty Molecular Biology

Date 2008 Jun 28

PMID 18583988

Citations 159

Authors

J Ross Chapman

Stephen P Jackson

Affiliations

Soon will be listed here.

Abstract

Mammalian cells respond to DNA double-strand breaks (DSBs) by recruiting DNA repair and cell-cycle checkpoint proteins to such sites. Central to these DNA damage response (DDR) events is the DNA damage mediator protein MDC1. MDC1 interacts with several DDR proteins, including the MRE11-RAD50-NBS1 (MRN) complex. Here, we show that MDC1 is phosphorylated on a cluster of conserved repeat motifs by casein kinase 2 (CK2). Moreover, we establish that this phosphorylation of MDC1 promotes direct, phosphorylation-dependent interactions with NBS1 in a manner that requires the closely apposed FHA and twin BRCT domains in the amino terminus of NBS1. Finally, we show that these CK2-targeted motifs in MDC1 are required to mediate NBS1 association with chromatin-flanking sites of unrepaired DSBs. These findings provide a molecular explanation for the MDC1-MRN interaction and yield insights into how MDC1 coordinates the focal assembly and activation of several DDR factors in response to DNA damage.

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References

Xie A, Hartlerode A, Stucki M, Odate S, Puget N, Kwok A . Distinct roles of chromatin-associated proteins MDC1 and 53BP1 in mammalian double-strand break repair. Mol Cell. 2007; 28(6):1045-57. PMC: 2275782. DOI: 10.1016/j.molcel.2007.12.005. View

Yu X, Chini C, He M, Mer G, Chen J . The BRCT domain is a phospho-protein binding domain. Science. 2003; 302(5645):639-42. DOI: 10.1126/science.1088753. View

Zhang D, Zaugg K, Mak T, Elledge S . A role for the deubiquitinating enzyme USP28 in control of the DNA-damage response. Cell. 2006; 126(3):529-42. DOI: 10.1016/j.cell.2006.06.039. View

Kastan M, Bartek J . Cell-cycle checkpoints and cancer. Nature. 2004; 432(7015):316-23. DOI: 10.1038/nature03097. View

Huen M, Grant R, Manke I, Minn K, Yu X, Yaffe M . RNF8 transduces the DNA-damage signal via histone ubiquitylation and checkpoint protein assembly. Cell. 2007; 131(5):901-14. PMC: 2149842. DOI: 10.1016/j.cell.2007.09.041. View

Iles N, Rulten S, El-Khamisy S, Caldecott K . APLF (C2orf13) is a novel human protein involved in the cellular response to chromosomal DNA strand breaks. Mol Cell Biol. 2007; 27(10):3793-803. PMC: 1900005. DOI: 10.1128/MCB.02269-06. View

Clements P, Breslin C, Deeks E, Byrd P, Ju L, Bieganowski P . The ataxia-oculomotor apraxia 1 gene product has a role distinct from ATM and interacts with the DNA strand break repair proteins XRCC1 and XRCC4. DNA Repair (Amst). 2004; 3(11):1493-502. DOI: 10.1016/j.dnarep.2004.06.017. View

Stucki M, Clapperton J, Mohammad D, Yaffe M, Smerdon S, Jackson S . MDC1 directly binds phosphorylated histone H2AX to regulate cellular responses to DNA double-strand breaks. Cell. 2005; 123(7):1213-26. DOI: 10.1016/j.cell.2005.09.038. View

Lou Z, Minter-Dykhouse K, Wu X, Chen J . MDC1 is coupled to activated CHK2 in mammalian DNA damage response pathways. Nature. 2003; 421(6926):957-61. DOI: 10.1038/nature01447. View

10.

Kolas N, Chapman J, Nakada S, Ylanko J, Chahwan R, Sweeney F . Orchestration of the DNA-damage response by the RNF8 ubiquitin ligase. Science. 2007; 318(5856):1637-40. PMC: 2430610. DOI: 10.1126/science.1150034. View

11.

Stucki M, Jackson S . gammaH2AX and MDC1: anchoring the DNA-damage-response machinery to broken chromosomes. DNA Repair (Amst). 2006; 5(5):534-43. DOI: 10.1016/j.dnarep.2006.01.012. View

12.

Mailand N, Bekker-Jensen S, Faustrup H, Melander F, Bartek J, Lukas C . RNF8 ubiquitylates histones at DNA double-strand breaks and promotes assembly of repair proteins. Cell. 2007; 131(5):887-900. DOI: 10.1016/j.cell.2007.09.040. View

13.

Olsen J, Blagoev B, Gnad F, Macek B, Kumar C, Mortensen P . Global, in vivo, and site-specific phosphorylation dynamics in signaling networks. Cell. 2006; 127(3):635-48. DOI: 10.1016/j.cell.2006.09.026. View

14.

Manke I, Lowery D, Nguyen A, Yaffe M . BRCT repeats as phosphopeptide-binding modules involved in protein targeting. Science. 2003; 302(5645):636-9. DOI: 10.1126/science.1088877. View

15.

Goldberg M, Stucki M, Falck J, DAmours D, Rahman D, Pappin D . MDC1 is required for the intra-S-phase DNA damage checkpoint. Nature. 2003; 421(6926):952-6. DOI: 10.1038/nature01445. View

16.

Durocher D, Taylor I, Sarbassova D, Haire L, Westcott S, Jackson S . The molecular basis of FHA domain:phosphopeptide binding specificity and implications for phospho-dependent signaling mechanisms. Mol Cell. 2000; 6(5):1169-82. DOI: 10.1016/s1097-2765(00)00114-3. View

17.

Becker E, Meyer V, Madaoui H, Guerois R . Detection of a tandem BRCT in Nbs1 and Xrs2 with functional implications in the DNA damage response. Bioinformatics. 2006; 22(11):1289-92. DOI: 10.1093/bioinformatics/btl075. View

18.

Stewart G, Wang B, Bignell C, Taylor A, Elledge S . MDC1 is a mediator of the mammalian DNA damage checkpoint. Nature. 2003; 421(6926):961-6. DOI: 10.1038/nature01446. View

19.

Beausoleil S, Jedrychowski M, Schwartz D, Elias J, Villen J, Li J . Large-scale characterization of HeLa cell nuclear phosphoproteins. Proc Natl Acad Sci U S A. 2004; 101(33):12130-5. PMC: 514446. DOI: 10.1073/pnas.0404720101. View

20.

Fernandez-Capetillo O, Lee A, Nussenzweig M, Nussenzweig A . H2AX: the histone guardian of the genome. DNA Repair (Amst). 2004; 3(8-9):959-67. DOI: 10.1016/j.dnarep.2004.03.024. View