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PMID: 18001825 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

RNF8 transduces the DNA-damage signal via histone ubiquitylation and checkpoint protein assembly.

Cell ·Vol. 131 ·No. 5 ·2007-11-30 ·Pages 901-14

Huen MS, Grant R, Manke I, Minn K, Yu X, Yaffe MB, Chen J

Abstract

DNA-damage signaling utilizes a multitude of posttranslational modifiers as molecular switches to regulate cell-cycle checkpoints, DNA repair, cellular senescence, and apoptosis. Here we show that RNF8, a FHA/RING domain-containing protein, plays a critical role in the early DNA-damage response. We have solved the X-ray crystal structure of the FHA domain structure at 1.35 A. We have shown that RNF8 facilitates the accumulation of checkpoint mediator proteins BRCA1 and 53BP1 to the damaged chromatin, on one hand through the phospho-dependent FHA domain-mediated binding of RNF8 to MDC1, on the other hand via its role in ubiquitylating H2AX and possibly other substrates at damage sites. Moreover, RNF8-depleted cells displayed a defective G2/M checkpoint and increased IR sensitivity. Together, our study implicates RNF8 as a novel DNA-damage-responsive protein that integrates protein phosphorylation and ubiquitylation signaling and plays a critical role in the cellular response to genotoxic stress.

MeSH Terms
Adaptor Proteins, Signal Transducing Amino Acid Motifs BRCA1 Protein/metabolism Binding Sites Cell Cycle/radiation effects Cell Cycle Proteins/metabolism Cells, Cultured DNA Damage/physiology DNA-Binding Proteins/chemistry,metabolism,physiology Histones/metabolism,physiology Humans Intracellular Signaling Peptides and Proteins/metabolism Models, Biological Models, Molecular Nuclear Proteins/physiology Phosphorylation Protein Binding Protein Structure, Tertiary Signal Transduction Trans-Activators/physiology Tumor Suppressor p53-Binding Protein 1 Ubiquitin-Protein Ligases Ubiquitination/physiology
Chemicals
Adaptor Proteins, Signal Transducing BRCA1 Protein BRCA1 protein, human Cell Cycle Proteins DNA-Binding Proteins H2AX protein, human Histones Intracellular Signaling Peptides and Proteins MDC1 protein, human Nuclear Proteins RNF8 protein, human TP53BP1 protein, human Trans-Activators Tumor Suppressor p53-Binding Protein 1 Ubiquitin-Protein Ligases
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Huen Michael S Y
Department of Therapeutic Radiology, Yale University School of Medicine, New Haven, CT 06520, USA.
Grant Robert
Manke Isaac
Minn Kay
Yu Xiaochun
Yaffe Michael B
Chen Junjie
References (49)
49 references, click to expand
  1. Rad53 FHA domain associated with phosphorylated Rad9 in the DNA damage checkpoint.
    Science. 1998 Jul 10;281(5374):272-4 PMID: 9657725
  2. N-Terminally extended human ubiquitin-conjugating enzymes (E2s) mediate the ubiquitination of RING-finger proteins, ARA54 and RNF8.
    Eur J Biochem. 2001 May;268(9):2725-32 PMID: 11322894
  3. BRCA1 : BARD1 induces the formation of conjugated ubiquitin structures, dependent on K6 of ubiquitin, in cells during DNA replication and repair.
    Hum Mol Genet. 2004 Apr 15;13(8):807-17 PMID: 14976165
  4. Control of spontaneous and damage-induced mutagenesis by SUMO and ubiquitin conjugation.
    Nature. 2003 Sep 11;425(6954):188-91 PMID: 12968183
  5. The FHA domain mediates phosphoprotein interactions.
    J Cell Sci. 2000 Dec;113 Pt 23:4143-9 PMID: 11069759
  6. Histone H2B monoubiquitination functions cooperatively with FACT to regulate elongation by RNA polymerase II.
    Cell. 2006 May 19;125(4):703-17 PMID: 16713563
  7. Accumulation of checkpoint protein 53BP1 at DNA breaks involves its binding to phosphorylated histone H2AX.
    J Biol Chem. 2003 May 30;278(22):19579-82 PMID: 12697768
  8. BRCT repeats as phosphopeptide-binding modules involved in protein targeting.
    Science. 2003 Oct 24;302(5645):636-9 PMID: 14576432
  9. Convergence of the fanconi anemia and ataxia telangiectasia signaling pathways.
    Cell. 2002 May 17;109(4):459-72 PMID: 12086603
  10. H2AX haploinsufficiency modifies genomic stability and tumor susceptibility.
    Cell. 2003 Aug 8;114(3):371-383 PMID: 12914701
  11. The BRCT domain is a phospho-protein binding domain.
    Science. 2003 Oct 24;302(5645):639-42 PMID: 14576433
  12. DNA damage-induced G2-M checkpoint activation by histone H2AX and 53BP1.
    Nat Cell Biol. 2002 Dec;4(12):993-7 PMID: 12447390
  13. Abraxas and RAP80 form a BRCA1 protein complex required for the DNA damage response.
    Science. 2007 May 25;316(5828):1194-8 PMID: 17525340
  14. Noncanonical MMS2-encoded ubiquitin-conjugating enzyme functions in assembly of novel polyubiquitin chains for DNA repair.
    Cell. 1999 Mar 5;96(5):645-53 PMID: 10089880
  15. MDC1 is required for the intra-S-phase DNA damage checkpoint.
    Nature. 2003 Feb 27;421(6926):952-6 PMID: 12607003
  16. The molecular basis of FHA domain:phosphopeptide binding specificity and implications for phospho-dependent signaling mechanisms.
    Mol Cell. 2000 Nov;6(5):1169-82 PMID: 11106755
  17. A critical role for histone H2AX in recruitment of repair factors to nuclear foci after DNA damage.
    Curr Biol. 2000 Jul 27-Aug 10;10(15):886-95 PMID: 10959836
  18. Histone H3 and H4 ubiquitylation by the CUL4-DDB-ROC1 ubiquitin ligase facilitates cellular response to DNA damage.
    Mol Cell. 2006 May 5;22(3):383-94 PMID: 16678110
  19. ATM and ATR substrate analysis reveals extensive protein networks responsive to DNA damage.
    Science. 2007 May 25;316(5828):1160-6 PMID: 17525332
  20. The RING finger protein, RNF8, interacts with retinoid X receptor alpha and enhances its transcription-stimulating activity.
    J Biol Chem. 2004 Apr 30;279(18):18926-34 PMID: 14981089
  21. The DDB1-CUL4ADDB2 ubiquitin ligase is deficient in xeroderma pigmentosum group E and targets histone H2A at UV-damaged DNA sites.
    Proc Natl Acad Sci U S A. 2006 Feb 21;103(8):2588-93 PMID: 16473935
  22. RAP80 targets BRCA1 to specific ubiquitin structures at DNA damage sites.
    Science. 2007 May 25;316(5828):1198-202 PMID: 17525341
  23. Interaction of the Fanconi anemia proteins and BRCA1 in a common pathway.
    Mol Cell. 2001 Feb;7(2):249-62 PMID: 11239454
  24. Structural basis for the methylation state-specific recognition of histone H4-K20 by 53BP1 and Crb2 in DNA repair.
    Cell. 2006 Dec 29;127(7):1361-73 PMID: 17190600
  25. BRCA1 ubiquitinates its phosphorylation-dependent binding partner CtIP.
    Genes Dev. 2006 Jul 1;20(13):1721-6 PMID: 16818604
  26. DNA damage response as a candidate anti-cancer barrier in early human tumorigenesis.
    Nature. 2005 Apr 14;434(7035):864-70 PMID: 15829956
  27. RAD6-dependent DNA repair is linked to modification of PCNA by ubiquitin and SUMO.
    Nature. 2002 Sep 12;419(6903):135-41 PMID: 12226657
  28. Structural and functional versatility of the FHA domain in DNA-damage signaling by the tumor suppressor kinase Chk2.
    Mol Cell. 2002 May;9(5):1045-54 PMID: 12049740
  29. Histone H2AX: a dosage-dependent suppressor of oncogenic translocations and tumors.
    Cell. 2003 Aug 8;114(3):359-70 PMID: 12914700
  30. The RING finger protein RNF8 recruits UBC13 for lysine 63-based self polyubiquitylation.
    J Cell Biochem. 2006 Feb 15;97(3):572-82 PMID: 16215985
  31. A conserved pathway to activate BRCA1-dependent ubiquitylation at DNA damage sites.
    EMBO J. 2006 May 17;25(10):2178-88 PMID: 16628214
  32. MDC1 directly binds phosphorylated histone H2AX to regulate cellular responses to DNA double-strand breaks.
    Cell. 2005 Dec 29;123(7):1213-26 PMID: 16377563
  33. Regulation of DNA repair by ubiquitylation.
    Nat Rev Mol Cell Biol. 2006 May;7(5):323-34 PMID: 16633336
  34. Chromatin remodelling at a DNA double-strand break site in Saccharomyces cerevisiae.
    Nature. 2005 Nov 17;438(7066):379-83 PMID: 16292314
  35. MDC1 is coupled to activated CHK2 in mammalian DNA damage response pathways.
    Nature. 2003 Feb 27;421(6926):957-61 PMID: 12607004
  36. MDC1 is a mediator of the mammalian DNA damage checkpoint.
    Nature. 2003 Feb 27;421(6926):961-6 PMID: 12607005
  37. A FancD2-monoubiquitin fusion reveals hidden functions of Fanconi anemia core complex in DNA repair.
    Mol Cell. 2005 Sep 16;19(6):841-7 PMID: 16168378
  38. Ubiquitin-binding protein RAP80 mediates BRCA1-dependent DNA damage response.
    Science. 2007 May 25;316(5828):1202-5 PMID: 17525342
  39. HKL-3000: the integration of data reduction and structure solution--from diffraction images to an initial model in minutes.
    Acta Crystallogr D Biol Crystallogr. 2006 Aug;62(Pt 8):859-66 PMID: 16855301
  40. MDC1 maintains genomic stability by participating in the amplification of ATM-dependent DNA damage signals.
    Mol Cell. 2006 Jan 20;21(2):187-200 PMID: 16427009
  41. Methylated lysine 79 of histone H3 targets 53BP1 to DNA double-strand breaks.
    Nature. 2004 Nov 18;432(7015):406-11 PMID: 15525939
  42. Histone acetylation by Trrap-Tip60 modulates loading of repair proteins and repair of DNA double-strand breaks.
    Nat Cell Biol. 2006 Jan;8(1):91-9 PMID: 16341205
  43. Likelihood-enhanced fast rotation functions.
    Acta Crystallogr D Biol Crystallogr. 2004 Mar;60(Pt 3):432-8 PMID: 14993666
  44. A critical role for the ubiquitin-conjugating enzyme Ubc13 in initiating homologous recombination.
    Mol Cell. 2007 Mar 9;25(5):663-75 PMID: 17349954
  45. Chfr is required for tumor suppression and Aurora A regulation.
    Nat Genet. 2005 Apr;37(4):401-6 PMID: 15793587
  46. Methylation of histone H4 lysine 20 controls recruitment of Crb2 to sites of DNA damage.
    Cell. 2004 Nov 24;119(5):603-14 PMID: 15550243
  47. DNA damage-induced cell cycle checkpoint control requires CtIP, a phosphorylation-dependent binding partner of BRCA1 C-terminal domains.
    Mol Cell Biol. 2004 Nov;24(21):9478-86 PMID: 15485915
  48. Activation of the DNA damage checkpoint and genomic instability in human precancerous lesions.
    Nature. 2005 Apr 14;434(7035):907-13 PMID: 15829965
  49. 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
Article Info
Journal
Cell
Abbr.
Cell
ISSN
0092-8674
DOI
10.1016/j.cell.2007.09.041
Published
2007-11-30
Epub
2007-00-20
Pages
901-14
Language
English
Region
United States
NLM ID
0413066
PMCID
PMC2149842
Subset
IM
Grants
NCI NIH HHS · R01 CA089239-08 · United States
NCI NIH HHS · CA89239 · United States
NCI NIH HHS · R01 CA100109 · United States
NIGMS NIH HHS · R01 GM060594 · United States
NCI NIH HHS · R01 CA089239 · United States
NCI NIH HHS · P50 CA116201 · United States
NCI NIH HHS · CA100109 · United States
NCI NIH HHS · CA92312 · United States
NIGMS NIH HHS · GM 60594 · United States
NCI NIH HHS · R01 CA092312-08 · United States
NCI NIH HHS · R01 CA092312 · United States
NCI NIH HHS · R01 CA100109-06 · United States
External Links
PubMed source DOI: 10.1016/j.cell.2007.09.041 PMC Full Text View journal details
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