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PMID: 15201865 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Mdc1 couples DNA double-strand break recognition by Nbs1 with its H2AX-dependent chromatin retention.

The EMBO journal ·Vol. 23 ·No. 13 ·2004-07-07 ·Pages 2674-83

Lukas C, Melander F, Stucki M, Falck J, Bekker-Jensen S, Goldberg M, Lerenthal Y, Jackson SP, Bartek J, Lukas J

Abstract

Mdc1/NFBD1 controls cellular responses to DNA damage, in part via interacting with the Mre11-Rad50-Nbs1 complex that is involved in the recognition, signalling, and repair of DNA double-strand breaks (DSBs). Here, we show that in live human cells, the transient interaction of Nbs1 with DSBs and its phosphorylation by ATM are Mdc1-independent. However, ablation of Mdc1 by siRNA or mutation of the Nbs1's FHA domain required for Mdc1 binding reduced the affinity of Nbs1 for DSB-flanking chromatin and caused aberrant pan-nuclear dispersal of Nbs1. This occurred despite normal phosphorylation of H2AX, indicating that lack of Mdc1 does not impair this DSB-induced chromatin change, but rather precludes the sustained engagement of Nbs1 with these regions. Mdc1 (but not Nbs1) became partially immobilized to chromatin after DSB generation, and siRNA-mediated depletion of H2AX prevented such relocalization of Mdc1 and uncoupled Nbs1 from DSB-flanking chromatin. Our data suggest that Mdc1 functions as an H2AX-dependent interaction platform enabling a switch from transient, Mdc1-independent recruitment of Nbs1 to DSBs towards sustained, Mdc1-dependent interactions with the surrounding chromosomal microenvironment.

MeSH Terms
Adaptor Proteins, Signal Transducing Antibodies, Monoclonal/metabolism Cell Cycle Proteins/chemistry,metabolism Cell Line, Tumor Cell Nucleus/metabolism Chromatin/metabolism DNA Damage DNA-Binding Proteins/metabolism Fluorescent Antibody Technique Fluorescent Dyes Histones/metabolism Humans Hydrazines Microscopy, Confocal Nuclear Proteins/chemistry,metabolism Osteosarcoma/pathology Phosphoproteins/metabolism Protein Structure, Tertiary RNA Interference RNA, Small Interfering/metabolism Trans-Activators/metabolism
Chemicals
Adaptor Proteins, Signal Transducing Alexa 488 hydrazide Antibodies, Monoclonal Cell Cycle Proteins Chromatin DNA-Binding Proteins Fluorescent Dyes H2AX protein, human Histones Hydrazines MDC1 protein, human NBN protein, human Nuclear Proteins Phosphoproteins RNA, Small Interfering Trans-Activators
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Lukas Claudia
Danish Cancer Society, Institute of Cancer Biology, Copenhagen, Denmark. [email protected]
Melander Fredrik
Stucki Manuel
Falck Jacob
Bekker-Jensen Simon
Goldberg Michal
Lerenthal Yaniv
Jackson Stephen P
Bartek Jiri
Lukas Jiri
References (37)
37 references, click to expand
  1. Substrate specificities and identification of putative substrates of ATM kinase family members.
    J Biol Chem. 1999 Dec 31;274(53):37538-43 PMID: 10608806
  2. The Nijmegen breakage syndrome protein is essential for Mre11 phosphorylation upon DNA damage.
    J Biol Chem. 1999 Jul 9;274(28):19513-6 PMID: 10391882
  3. ATM-dependent phosphorylation of nibrin in response to radiation exposure.
    Nat Genet. 2000 May;25(1):115-9 PMID: 10802669
  4. Functional link between ataxia-telangiectasia and Nijmegen breakage syndrome gene products.
    Nature. 2000 May 25;405(6785):473-7 PMID: 10839544
  5. NFBD1/KIAA0170 is a novel nuclear transcriptional transactivator with BRCT domain.
    DNA Cell Biol. 2000 Aug;19(8):475-85 PMID: 10975465
  6. Chromatin association of human origin recognition complex, cdc6, and minichromosome maintenance proteins during the cell cycle: assembly of prereplication complexes in late mitosis.
    Mol Cell Biol. 2000 Nov;20(22):8602-12 PMID: 11046155
  7. DNA damage-dependent nuclear dynamics of the Mre11 complex.
    Mol Cell Biol. 2001 Jan;21(1):281-8 PMID: 11113202
  8. Distinct functional domains of nibrin mediate Mre11 binding, focus formation, and nuclear localization.
    Mol Cell Biol. 2001 Mar;21(6):2184-91 PMID: 11238951
  9. The DNA damage-dependent intra-S phase checkpoint is regulated by parallel pathways.
    Nat Genet. 2002 Mar;30(3):290-4 PMID: 11850621
  10. Nuclear dynamics of RAD52 group homologous recombination proteins in response to DNA damage.
    EMBO J. 2002 Apr 15;21(8):2030-7 PMID: 11953322
  11. A system for stable expression of short interfering RNAs in mammalian cells.
    Science. 2002 Apr 19;296(5567):550-3 PMID: 11910072
  12. The Mre11 complex: at the crossroads of dna repair and checkpoint signalling.
    Nat Rev Mol Cell Biol. 2002 May;3(5):317-27 PMID: 11988766
  13. NBS1 localizes to gamma-H2AX foci through interaction with the FHA/BRCT domain.
    Curr Biol. 2002 Oct 29;12(21):1846-51 PMID: 12419185
  14. Nijmegen breakage syndrome gene, NBS1, and molecular links to factors for genome stability.
    Oncogene. 2002 Dec 16;21(58):8967-80 PMID: 12483513
  15. DNA damage activates ATM through intermolecular autophosphorylation and dimer dissociation.
    Nature. 2003 Jan 30;421(6922):499-506 PMID: 12556884
  16. NFBD1, a novel nuclear protein with signature motifs of FHA and BRCT, and an internal 41-amino acid repeat sequence, is an early participant in DNA damage response.
    J Biol Chem. 2003 Feb 21;278(8):6323-9 PMID: 12475977
  17. MDC1 is required for the intra-S-phase DNA damage checkpoint.
    Nature. 2003 Feb 27;421(6926):952-6 PMID: 12607003
  18. MDC1 is a mediator of the mammalian DNA damage checkpoint.
    Nature. 2003 Feb 27;421(6926):961-6 PMID: 12607005
  19. NFBD1/KIAA0170 is a chromatin-associated protein involved in DNA damage signaling pathways.
    J Biol Chem. 2003 Mar 7;278(10):8795-803 PMID: 12499369
  20. ATM and related protein kinases: safeguarding genome integrity.
    Nat Rev Cancer. 2003 Mar;3(3):155-68 PMID: 12612651
  21. NFBD1, like 53BP1, is an early and redundant transducer mediating Chk2 phosphorylation in response to DNA damage.
    J Biol Chem. 2003 Mar 14;278(11):8873-6 PMID: 12551934
  22. Distinct spatiotemporal dynamics of mammalian checkpoint regulators induced by DNA damage.
    Nat Cell Biol. 2003 Mar;5(3):255-60 PMID: 12598907
  23. Mediator of DNA damage checkpoint protein 1 regulates BRCA1 localization and phosphorylation in DNA damage checkpoint control.
    J Biol Chem. 2003 Apr 18;278(16):13599-602 PMID: 12611903
  24. Nibrin forkhead-associated domain and breast cancer C-terminal domain are both required for nuclear focus formation and phosphorylation.
    J Biol Chem. 2003 Jun 13;278(24):21944-51 PMID: 12679336
  25. Histone H2AX phosphorylation is dispensable for the initial recognition of DNA breaks.
    Nat Cell Biol. 2003 Jul;5(7):675-9 PMID: 12792649
  26. Distinct functions of Nijmegen breakage syndrome in ataxia telangiectasia mutated-dependent responses to DNA damage.
    Mol Cancer Res. 2003 Jul;1(9):674-81 PMID: 12861053
  27. The cellular response to DNA double-strand breaks: defining the sensors and mediators.
    Trends Cell Biol. 2003 Sep;13(9):458-62 PMID: 12946624
  28. Megabase chromatin domains involved in DNA double-strand breaks in vivo.
    J Cell Biol. 1999 Sep 6;146(5):905-16 PMID: 10477747
  29. Requirement of the MRN complex for ATM activation by DNA damage.
    EMBO J. 2003 Oct 15;22(20):5612-21 PMID: 14532133
  30. The MRN complex: coordinating and mediating the response to broken chromosomes.
    EMBO Rep. 2003 Sep;4(9):844-9 PMID: 12949583
  31. NFBD1/MDC1 regulates ionizing radiation-induced focus formation by DNA checkpoint signaling and repair factors.
    FASEB J. 2003 Oct;17(13):1842-8 PMID: 14519663
  32. The Mre11 complex is required for ATM activation and the G2/M checkpoint.
    EMBO J. 2003 Dec 15;22(24):6610-20 PMID: 14657032
  33. 53BP1 and NFBD1/MDC1-Nbs1 function in parallel interacting pathways activating ataxia-telangiectasia mutated (ATM) in response to DNA damage.
    Cancer Res. 2003 Dec 15;63(24):8586-91 PMID: 14695167
  34. Distinct functional domains of Nbs1 modulate the timing and magnitude of ATM activation after low doses of ionizing radiation.
    Oncogene. 2004 Apr 15;23(17):3122-7 PMID: 15048089
  35. Radiosensitivity in ataxia-telangiectasia: a new explanation.
    Proc Natl Acad Sci U S A. 1980 Dec;77(12):7315-7 PMID: 6938978
  36. A new method for introducing double-strand breaks into cellular DNA.
    Radiat Res. 1993 May;134(2):160-9 PMID: 7683818
  37. ATM phosphorylates p95/nbs1 in an S-phase checkpoint pathway.
    Nature. 2000 Apr 6;404(6778):613-7 PMID: 10766245
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1038/sj.emboj.7600269
Email
[email protected]
Published
2004-07-07
Epub
2004-00-17
Pages
2674-83
Language
English
Region
England
NLM ID
8208664
PMCID
PMC449779
Subset
IM
External Links
PubMed source DOI: 10.1038/sj.emboj.7600269 PMC Full Text View journal details
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