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PMID: 19221393 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

MRN complex function in the repair of chromosomal Rag-mediated DNA double-strand breaks.

The Journal of experimental medicine ·Vol. 206 ·No. 3 ·2009-03-16 ·Pages 669-79

Helmink BA, Bredemeyer AL, Lee BS, Huang CY, Sharma GG, Walker LM, Bednarski JJ, Lee WL, Pandita TK, Bassing CH, Sleckman BP

Abstract

The Mre11-Rad50-Nbs1 (MRN) complex functions in the repair of DNA double-strand breaks (DSBs) by homologous recombination (HR) at postreplicative stages of the cell cycle. During HR, the MRN complex functions directly in the repair of DNA DSBs and in the initiation of DSB responses through activation of the ataxia telangiectasia-mutated (ATM) serine-threonine kinase. Whether MRN functions in DNA damage responses before DNA replication in G0/G1 phase cells has been less clear. In developing G1-phase lymphocytes, DNA DSBs are generated by the Rag endonuclease and repaired during the assembly of antigen receptor genes by the process of V(D)J recombination. Mice and humans deficient in MRN function exhibit lymphoid phenotypes that are suggestive of defects in V(D)J recombination. We show that during V(D)J recombination, MRN deficiency leads to the aberrant joining of Rag DSBs and to the accumulation of unrepaired coding ends, thus establishing a functional role for MRN in the repair of Rag-mediated DNA DSBs. Moreover, these defects in V(D)J recombination are remarkably similar to those observed in ATM-deficient lymphocytes, suggesting that ATM and MRN function in the same DNA DSB response pathways during lymphocyte antigen receptor gene assembly.

MeSH Terms
ATP-Binding Cassette Transporters/metabolism Acid Anhydride Hydrolases Animals Ataxia Telangiectasia Mutated Proteins Cell Cycle Proteins/antagonists & inhibitors,metabolism Chromosomes, Mammalian/metabolism DNA Breaks, Double-Stranded DNA Repair DNA Repair Enzymes/deficiency,metabolism DNA-Binding Proteins/antagonists & inhibitors,deficiency,metabolism Enzyme Activation Homeodomain Proteins/metabolism Humans MRE11 Homologue Protein Mice Nuclear Proteins/deficiency,metabolism Oncogene Proteins v-abl/metabolism Precursor Cells, B-Lymphoid/enzymology Protein Serine-Threonine Kinases/antagonists & inhibitors Recombination, Genetic/genetics Retroviridae Thymus Gland/cytology Tumor Suppressor Proteins/antagonists & inhibitors VDJ Exons/genetics
Chemicals
ATP-Binding Cassette Transporters Cell Cycle Proteins DNA-Binding Proteins Homeodomain Proteins Mre11a protein, mouse Nijmegen breakage syndrome 1 protein, mouse Nuclear Proteins Oncogene Proteins v-abl Tumor Suppressor Proteins RAG-1 protein ATM protein, human Ataxia Telangiectasia Mutated Proteins Atm protein, mouse Protein Serine-Threonine Kinases MRE11 Homologue Protein Acid Anhydride Hydrolases Rad50 protein, mouse DNA Repair Enzymes
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Helmink Beth A
Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110, USA.
Bredemeyer Andrea L
Lee Baeck-Seung
Huang Ching-Yu
Sharma Girdhar G
Walker Laura M
Bednarski Jeffrey J
Lee Wan-Ling
Pandita Tej K
Bassing Craig H
Sleckman Barry P
References (73)
73 references, click to expand
  1. Abnormal development of Purkinje cells and lymphocytes in Atm mutant mice.
    Proc Natl Acad Sci U S A. 2000 Mar 28;97(7):3336-41 PMID: 10716718
  2. ATM-dependent phosphorylation of nibrin in response to radiation exposure.
    Nat Genet. 2000 May;25(1):115-9 PMID: 10802669
  3. In situ visualization of DNA double-strand break repair in human fibroblasts.
    Science. 1998 Apr 24;280(5363):590-2 PMID: 9554850
  4. ATM phosphorylation of Nijmegen breakage syndrome protein is required in a DNA damage response.
    Nature. 2000 May 25;405(6785):477-82 PMID: 10839545
  5. The carboxy terminus of NBS1 is required for induction of apoptosis by the MRE11 complex.
    Nature. 2007 May 10;447(7141):218-21 PMID: 17429352
  6. Structure of the Rad50 x Mre11 DNA repair complex from Saccharomyces cerevisiae by electron microscopy.
    J Biol Chem. 2001 Oct 5;276(40):37027-33 PMID: 11470800
  7. Interplay between human DNA repair proteins at a unique double-strand break in vivo.
    EMBO J. 2006 Jan 11;25(1):222-31 PMID: 16395335
  8. Nijmegen breakage syndrome: clinical manifestation of defective response to DNA double-strand breaks.
    DNA Repair (Amst). 2004 Aug-Sep;3(8-9):1207-17 PMID: 15279809
  9. Lymphocyte-specific compensation for XLF/cernunnos end-joining functions in V(D)J recombination.
    Mol Cell. 2008 Sep 5;31(5):631-40 PMID: 18775323
  10. Response to RAG-mediated VDJ cleavage by NBS1 and gamma-H2AX.
    Science. 2000 Dec 8;290(5498):1962-5 PMID: 11110662
  11. Aberrant V(D)J recombination in ataxia telangiectasia mutated-deficient lymphocytes is dependent on nonhomologous DNA end joining.
    J Immunol. 2008 Aug 15;181(4):2620-5 PMID: 18684952
  12. Mammalian Rad9 plays a role in telomere stability, S- and G2-phase-specific cell survival, and homologous recombinational repair.
    Mol Cell Biol. 2006 Mar;26(5):1850-64 PMID: 16479004
  13. Disruption of mRad50 causes embryonic stem cell lethality, abnormal embryonic development, and sensitivity to ionizing radiation.
    Proc Natl Acad Sci U S A. 1999 Jun 22;96(13):7376-81 PMID: 10377422
  14. Targeted disruption of NBS1 reveals its roles in mouse development and DNA repair.
    EMBO J. 2002 Mar 15;21(6):1447-55 PMID: 11889050
  15. Somatic generation of antibody diversity.
    Nature. 1983 Apr 14;302(5909):575-81 PMID: 6300689
  16. ATM and ATR substrate analysis reveals extensive protein networks responsive to DNA damage.
    Science. 2007 May 25;316(5828):1160-6 PMID: 17525332
  17. Cell cycle and genetic requirements of two pathways of nonhomologous end-joining repair of double-strand breaks in Saccharomyces cerevisiae.
    Mol Cell Biol. 1996 May;16(5):2164-73 PMID: 8628283
  18. ATM prevents the persistence and propagation of chromosome breaks in lymphocytes.
    Cell. 2007 Jul 13;130(1):63-75 PMID: 17599403
  19. ATM deficiency disrupts Tcra locus integrity and the maturation of CD4+CD8+ thymocytes.
    Blood. 2007 Mar 1;109(5):1887-96 PMID: 17077325
  20. Revision of T cell receptor {alpha} chain genes is required for normal T lymphocyte development.
    Proc Natl Acad Sci U S A. 2005 Oct 4;102(40):14356-61 PMID: 16186502
  21. 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
  22. bcl-2 transgene inhibits T cell death and perturbs thymic self-censorship.
    Cell. 1991 Nov 29;67(5):889-99 PMID: 1959134
  23. DNA double-strand break repair: all's well that ends well.
    Annu Rev Genet. 2006;40:363-83 PMID: 16895466
  24. Direct activation of the ATM protein kinase by the Mre11/Rad50/Nbs1 complex.
    Science. 2004 Apr 2;304(5667):93-6 PMID: 15064416
  25. hMre11 and hRad50 nuclear foci are induced during the normal cellular response to DNA double-strand breaks.
    Mol Cell Biol. 1997 Oct;17(10):6087-96 PMID: 9315668
  26. Mesoscale conformational changes in the DNA-repair complex Rad50/Mre11/Nbs1 upon binding DNA.
    Nature. 2005 Sep 15;437(7057):440-3 PMID: 16163361
  27. Hyperthermia activates a subset of ataxia-telangiectasia mutated effectors independent of DNA strand breaks and heat shock protein 70 status.
    Cancer Res. 2007 Apr 1;67(7):3010-7 PMID: 17409407
  28. DNA double-strand breaks activate a multi-functional genetic program in developing lymphocytes.
    Nature. 2008 Dec 11;456(7223):819-23 PMID: 18849970
  29. Defective signal joint recombination in fanconi anemia fibroblasts reveals a role for Rad50 in V(D)J recombination.
    J Mol Biol. 2007 Jul 13;370(3):449-58 PMID: 17524422
  30. Interfaces between the detection, signaling, and repair of DNA damage.
    Science. 2002 Jul 26;297(5581):547-51 PMID: 12142523
  31. Conditional deletion of Nbs1 in murine cells reveals its role in branching repair pathways of DNA double-strand breaks.
    EMBO J. 2006 Nov 29;25(23):5527-38 PMID: 17082765
  32. Leaky Scid phenotype associated with defective V(D)J coding end processing in Artemis-deficient mice.
    Mol Cell. 2002 Dec;10(6):1379-90 PMID: 12504013
  33. Role of Nbs1 in the activation of the Atm kinase revealed in humanized mouse models.
    Nat Cell Biol. 2005 Jul;7(7):675-85 PMID: 15965469
  34. The RAG proteins and V(D)J recombination: complexes, ends, and transposition.
    Annu Rev Immunol. 2000;18:495-527 PMID: 10837067
  35. Chromatin relaxation in response to DNA double-strand breaks is modulated by a novel ATM- and KAP-1 dependent pathway.
    Nat Cell Biol. 2006 Aug;8(8):870-6 PMID: 16862143
  36. Distinct domains in Nbs1 regulate irradiation-induced checkpoints and apoptosis.
    J Exp Med. 2007 May 14;204(5):1003-11 PMID: 17485521
  37. Hairpin opening and overhang processing by an Artemis/DNA-dependent protein kinase complex in nonhomologous end joining and V(D)J recombination.
    Cell. 2002 Mar 22;108(6):781-94 PMID: 11955432
  38. Human Rad50/Mre11 is a flexible complex that can tether DNA ends.
    Mol Cell. 2001 Nov;8(5):1129-35 PMID: 11741547
  39. V(D)J rearrangement in Nijmegen breakage syndrome.
    Mol Immunol. 2000 Dec;37(18):1131-9 PMID: 11451418
  40. Atm-deficient mice: a paradigm of ataxia telangiectasia.
    Cell. 1996 Jul 12;86(1):159-71 PMID: 8689683
  41. ATM activation by DNA double-strand breaks through the Mre11-Rad50-Nbs1 complex.
    Science. 2005 Apr 22;308(5721):551-4 PMID: 15790808
  42. Novel functional requirements for non-homologous DNA end joining in Schizosaccharomyces pombe.
    EMBO J. 2001 Jan 15;20(1-2):210-21 PMID: 11226171
  43. Mre11 dimers coordinate DNA end bridging and nuclease processing in double-strand-break repair.
    Cell. 2008 Oct 3;135(1):97-109 PMID: 18854158
  44. Requirement of the MRN complex for ATM activation by DNA damage.
    EMBO J. 2003 Oct 15;22(20):5612-21 PMID: 14532133
  45. The DNA damage response: putting checkpoints in perspective.
    Nature. 2000 Nov 23;408(6811):433-9 PMID: 11100718
  46. The MRE11-RAD50-XRS2 complex, in addition to other non-homologous end-joining factors, is required for V(D)J joining in yeast.
    J Biol Chem. 2005 May 27;280(21):20247-52 PMID: 15757898
  47. Conditional gene targeted deletion by Cre recombinase demonstrates the requirement for the double-strand break repair Mre11 protein in murine embryonic stem cells.
    Nucleic Acids Res. 1997 Aug 1;25(15):2985-91 PMID: 9224597
  48. ATM stabilizes DNA double-strand-break complexes during V(D)J recombination.
    Nature. 2006 Jul 27;442(7101):466-70 PMID: 16799570
  49. V(D)J recombination: site-specific cleavage and repair.
    Mol Cells. 2000 Aug 31;10(4):367-74 PMID: 10987131
  50. Targeted disruption of the Nijmegen breakage syndrome gene NBS1 leads to early embryonic lethality in mice.
    Curr Biol. 2001 Jan 23;11(2):105-9 PMID: 11231126
  51. ATM and related protein kinases: safeguarding genome integrity.
    Nat Rev Cancer. 2003 Mar;3(3):155-68 PMID: 12612651
  52. The Mre11 complex and the metabolism of chromosome breaks: the importance of communicating and holding things together.
    DNA Repair (Amst). 2004 Aug-Sep;3(8-9):845-54 PMID: 15279769
  53. The role of the non-homologous end-joining pathway in lymphocyte development.
    Immunol Rev. 2004 Aug;200:115-31 PMID: 15242400
  54. The Rad50 zinc-hook is a structure joining Mre11 complexes in DNA recombination and repair.
    Nature. 2002 Aug 1;418(6897):562-6 PMID: 12152085
  55. Ataxia-telangiectasia-like disorder (ATLD)-its clinical presentation and molecular basis.
    DNA Repair (Amst). 2004 Aug-Sep;3(8-9):1219-25 PMID: 15279810
  56. Mre11 nuclease activity has essential roles in DNA repair and genomic stability distinct from ATM activation.
    Cell. 2008 Oct 3;135(1):85-96 PMID: 18854157
  57. Defects in coding joint formation in vivo in developing ATM-deficient B and T lymphocytes.
    J Exp Med. 2007 Jun 11;204(6):1371-81 PMID: 17502661
  58. Sensing of intermediates in V(D)J recombination by ATM.
    Genes Dev. 2002 Jan 15;16(2):159-64 PMID: 11799059
  59. A small molecule Abl kinase inhibitor induces differentiation of Abelson virus-transformed pre-B cell lines.
    Nat Immunol. 2003 Jan;4(1):31-7 PMID: 12469118
  60. Novel strand exchanges in V(D)J recombination.
    Cell. 1988 Dec 23;55(6):1099-107 PMID: 3144437
  61. The cell cycle and V(D)J recombination.
    Curr Top Microbiol Immunol. 1996;217:45-59 PMID: 8787617
  62. Components of the Ku-dependent non-homologous end-joining pathway are involved in telomeric length maintenance and telomeric silencing.
    EMBO J. 1998 Mar 16;17(6):1819-28 PMID: 9501103
  63. V(D)J recombination in ataxia telangiectasia, Bloom's syndrome, and a DNA ligase I-associated immunodeficiency disorder.
    J Biol Chem. 1993 Sep 25;268(27):20105-9 PMID: 8397200
  64. ATM deficiency impairs thymocyte maturation because of defective resolution of T cell receptor alpha locus coding end breaks.
    Proc Natl Acad Sci U S A. 2007 Apr 10;104(15):6323-8 PMID: 17405860
  65. Reconstitution of the mammalian DNA double-strand break end-joining reaction reveals a requirement for an Mre11/Rad50/NBS1-containing fraction.
    Nucleic Acids Res. 2002 Feb 1;30(3):667-74 PMID: 11809878
  66. A murine model of Nijmegen breakage syndrome.
    Curr Biol. 2002 Apr 16;12(8):648-53 PMID: 11967151
  67. ATM phosphorylates p95/nbs1 in an S-phase checkpoint pathway.
    Nature. 2000 Apr 6;404(6778):613-7 PMID: 10766245
  68. Checkpoint failure and chromosomal instability without lymphomagenesis in Mre11(ATLD1/ATLD1) mice.
    Mol Cell. 2003 Dec;12(6):1511-23 PMID: 14690604
  69. Normal V(D)J recombination in cells from patients with Nijmegen breakage syndrome.
    Mol Immunol. 2000 Oct;37(15):915-29 PMID: 11282395
  70. Repair of double-strand breaks by nonhomologous end joining in the absence of Mre11.
    J Cell Biol. 2005 Dec 5;171(5):765-71 PMID: 16330708
  71. Unusual immunoglobulin gene rearrangement leads to replacement of recombinational signal sequences.
    Science. 1988 Oct 14;242(4876):261-3 PMID: 3140378
  72. The Rad50 hook domain is a critical determinant of Mre11 complex functions.
    Nat Struct Mol Biol. 2005 May;12(5):403-7 PMID: 15852023
  73. The mechanism and regulation of chromosomal V(D)J recombination.
    Cell. 2002 Apr;109 Suppl:S45-55 PMID: 11983152
Article Info
Journal
The Journal of experimental medicine
Abbr.
J Exp Med
ISSN
1540-9538
DOI
10.1084/jem.20081326
Published
2009-03-16
Epub
2009-00-16
Pages
669-79
Language
English
Region
United States
NLM ID
2985109R
PMCID
PMC2699138
Subset
IM
Grants
NIAID NIH HHS · AI47829 · United States
NCI NIH HHS · R01 CA123232 · United States
NIAID NIH HHS · R01 AI074953 · United States
NCI NIH HHS · R01 CA129537 · United States
NICHD NIH HHS · 5 T32 HD007499 · United States
NIAID NIH HHS · AI074953 · United States
NIAID NIH HHS · R01 AI047829 · United States
NCI NIH HHS · CA123232 · United States
NCI NIH HHS · R01 CA125195 · United States
NICHD NIH HHS · T32 HD007499 · United States
NCI NIH HHS · CA129537 · United States
NCI NIH HHS · R01 CA125195-02 · United States
External Links
PubMed source DOI: 10.1084/jem.20081326 PMC Full Text View journal details
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