Home LiteratureArticle Details
PMID: 19748363 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Checkpoint signaling from a single DNA interstrand crosslink.

Molecular cell ·Vol. 35 ·No. 5 ·2009-09-11 ·Pages 704-15

Ben-Yehoyada M, Wang LC, Kozekov ID, Rizzo CJ, Gottesman ME, Gautier J

Abstract

DNA interstrand crosslinks (ICLs) are the most toxic lesions induced by chemotherapeutic agents such as mitomycin C and cisplatin. By covalently linking both DNA strands, ICLs prevent DNA melting, transcription, and replication. Studies on ICL signaling and repair have been limited, because these drugs generate additional DNA lesions that trigger checkpoint signaling. Here, we monitor sensing, signaling from, and repairing of a single site-specific ICL in cell-free extract derived from Xenopus eggs and in mammalian cells. Notably, we demonstrate that ICLs trigger a checkpoint response independently of origin-initiated DNA replication and uncoupling of DNA polymerase and DNA helicase. The Fanconi anemia pathway acts upstream of RPA-ATR-Chk1 to generate the ICL signal. The system also repairs ICLs in a reaction that involves extensive, error-free DNA synthesis. Repair occurs by both origin-dependent and origin-independent mechanisms. Our data suggest that cell sensitivity to crosslinking agents results from both checkpoint and DNA repair defects.

MeSH Terms
Alkylating Agents/pharmacology Animals Ataxia Telangiectasia Mutated Proteins Cell Cycle/genetics Cell Cycle Proteins/metabolism Cell Proliferation Checkpoint Kinase 1 DNA/biosynthesis,chemistry,metabolism DNA Damage DNA Helicases/metabolism DNA Repair DNA Replication DNA-Directed DNA Polymerase/metabolism Fanconi Anemia Complementation Group A Protein/metabolism Fanconi Anemia Complementation Group D2 Protein/metabolism HeLa Cells Humans Nucleic Acid Conformation Protein Kinases/metabolism Protein Serine-Threonine Kinases/metabolism Recombinant Proteins/metabolism Replication Origin Replication Protein A/metabolism Signal Transduction/genetics Time Factors Transfection Xenopus Proteins Xenopus laevis
Chemicals
Alkylating Agents Cell Cycle Proteins FANCA protein, human FANCD2 protein, human Fanconi Anemia Complementation Group A Protein Fanconi Anemia Complementation Group D2 Protein Recombinant Proteins Replication Protein A Xenopus Proteins DNA Protein Kinases ATR protein, human Ataxia Telangiectasia Mutated Proteins CHEK1 protein, human Checkpoint Kinase 1 Chek1 protein, Xenopus Protein Serine-Threonine Kinases DNA-Directed DNA Polymerase DNA Helicases
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Ben-Yehoyada Merav
Institute for Cancer Genetics, Department of Genetics and Development, Columbia University, New York, NY 10032, USA.
Wang Lily C
Kozekov Ivan D
Rizzo Carmelo J
Gottesman Max E
Gautier Jean
References (53)
53 references, click to expand
  1. Isolation and characterization of Xenopus ATM (X-ATM): expression, localization, and complex formation during oogenesis and early development.
    Oncogene. 1999 Nov 25;18(50):7070-9 PMID: 10597308
  2. Recombinant replication protein A: expression, complex formation, and functional characterization.
    J Biol Chem. 1994 Apr 15;269(15):11121-32 PMID: 8157639
  3. Fanconi anemia and DNA replication repair.
    DNA Repair (Amst). 2007 Jul 1;6(7):885-90 PMID: 17481966
  4. Emergence of a DNA-damage response network consisting of Fanconi anaemia and BRCA proteins.
    Nat Rev Genet. 2007 Oct;8(10):735-48 PMID: 17768402
  5. DNA interstrand crosslink repair during G1 involves nucleotide excision repair and DNA polymerase zeta.
    EMBO J. 2006 Mar 22;25(6):1285-94 PMID: 16482220
  6. Functional uncoupling of MCM helicase and DNA polymerase activities activates the ATR-dependent checkpoint.
    Genes Dev. 2005 May 1;19(9):1040-52 PMID: 15833913
  7. The ATPase activity of MCM2-7 is dispensable for pre-RC assembly but is required for DNA unwinding.
    EMBO J. 2005 Dec 21;24(24):4334-44 PMID: 16369567
  8. Regulated chromosomal DNA replication in the absence of a nucleus.
    Mol Cell. 1998 Mar;1(4):519-29 PMID: 9660936
  9. Mechanism of replication-coupled DNA interstrand crosslink repair.
    Cell. 2008 Sep 19;134(6):969-80 PMID: 18805090
  10. Repair of psoralen interstrand cross-links in Xenopus laevis egg extracts is highly mutagenic.
    Biochem Biophys Res Commun. 2005 Oct 14;336(1):69-75 PMID: 16125670
  11. S-phase-specific interaction of the Fanconi anemia protein, FANCD2, with BRCA1 and RAD51.
    Blood. 2002 Oct 1;100(7):2414-20 PMID: 12239151
  12. Repair of intermediate structures produced at DNA interstrand cross-links in Saccharomyces cerevisiae.
    Mol Cell Biol. 2000 May;20(10):3425-33 PMID: 10779332
  13. DNA cross-link-dependent RAD50/MRE11/NBS1 subnuclear assembly requires the Fanconi anemia C protein.
    Hum Mol Genet. 2002 Oct 1;11(21):2531-46 PMID: 12354779
  14. Chemosensitization to cisplatin by inhibitors of the Fanconi anemia/BRCA pathway.
    Mol Cancer Ther. 2006 Apr;5(4):952-61 PMID: 16648566
  15. The Fanconi anemia pathway and ubiquitin.
    BMC Biochem. 2007 Nov 22;8 Suppl 1:S10 PMID: 18047734
  16. FANCJ helicase defective in Fanconia anemia and breast cancer unwinds G-quadruplex DNA to defend genomic stability.
    Mol Cell Biol. 2008 Jun;28(12):4116-28 PMID: 18426915
  17. DNA interstrand cross-links induce futile repair synthesis in mammalian cell extracts.
    Mol Cell Biol. 2000 Apr;20(7):2446-54 PMID: 10713168
  18. Fanconi anemia (cross)linked to DNA repair.
    Cell. 2005 Dec 29;123(7):1191-8 PMID: 16377561
  19. Sensing DNA damage through ATRIP recognition of RPA-ssDNA complexes.
    Science. 2003 Jun 6;300(5625):1542-8 PMID: 12791985
  20. ATR and ATM regulate the timing of DNA replication origin firing.
    Nat Cell Biol. 2004 Jul;6(7):648-55 PMID: 15220931
  21. The fidelity of DNA synthesis by eukaryotic replicative and translesion synthesis polymerases.
    Cell Res. 2008 Jan;18(1):148-61 PMID: 18166979
  22. An ATR- and BRCA1-mediated Fanconi anemia pathway is required for activating the G2/M checkpoint and DNA damage repair upon rereplication.
    Mol Cell Biol. 2006 Jun;26(12):4601-11 PMID: 16738325
  23. A novel cell-free screen identifies a potent inhibitor of the Fanconi anemia pathway.
    Int J Cancer. 2009 Feb 15;124(4):783-92 PMID: 19048618
  24. Structural studies of an oligodeoxynucleotide containing a trimethylene interstrand cross-link in a 5'-(CpG) motif: model of a malondialdehyde cross-link.
    J Am Chem Soc. 2001 Feb 28;123(8):1730-9 PMID: 11456774
  25. Lack of sensitivity of primary Fanconi's anemia fibroblasts to UV and ionizing radiation.
    Radiat Res. 2004 Mar;161(3):318-25 PMID: 14982482
  26. The structure-specific endonuclease Mus81-Eme1 promotes conversion of interstrand DNA crosslinks into double-strands breaks.
    EMBO J. 2006 Oct 18;25(20):4921-32 PMID: 17036055
  27. Formation and repair of interstrand cross-links in DNA.
    Chem Rev. 2006 Feb;106(2):277-301 PMID: 16464006
  28. Deficiency in incisions produced by XPF at the site of a DNA interstrand cross-link in Fanconi anemia cells.
    Biochemistry. 2007 Dec 18;46(50):14359-68 PMID: 18020456
  29. Damage-resistant DNA synthesis in Fanconi anemia cells treated with a DNA cross-linking agent.
    Exp Cell Res. 2000 Nov 1;260(2):216-21 PMID: 11035916
  30. Fanconi anemia proteins stabilize replication forks.
    DNA Repair (Amst). 2008 Dec 1;7(12):1973-81 PMID: 18786657
  31. The structure-specific endonuclease Ercc1-Xpf is required to resolve DNA interstrand cross-link-induced double-strand breaks.
    Mol Cell Biol. 2004 Jul;24(13):5776-87 PMID: 15199134
  32. Characterization of a simian virus 40-transformed Fanconi anemia fibroblast cell line.
    Mutat Res. 1986 Sep;166(2):207-14 PMID: 3020399
  33. Replication protein A: a heterotrimeric, single-stranded DNA-binding protein required for eukaryotic DNA metabolism.
    Annu Rev Biochem. 1997;66:61-92 PMID: 9242902
  34. Isolation and characterization of pso mutants sensitive to photo-addition of psoralen derivatives in Saccharomyces cerevisiae.
    Genetics. 1980 Jun;95(2):273-88 PMID: 7009316
  35. Fanconi anemia proteins and the s phase checkpoint.
    Cell Cycle. 2004 Jun;3(6):698-700 PMID: 15136767
  36. NMR determination of the conformation of a trimethylene interstrand cross-link in an oligodeoxynucleotide duplex containing a 5'-d(GpC) motif.
    J Am Chem Soc. 2003 Jan 8;125(1):62-72 PMID: 12515507
  37. The Fanconi anemia core complex is required for efficient point mutagenesis and Rev1 foci assembly.
    DNA Repair (Amst). 2008 Jun 1;7(6):902-11 PMID: 18448394
  38. Multiple repair pathways mediate tolerance to chemotherapeutic cross-linking agents in vertebrate cells.
    Cancer Res. 2005 Dec 15;65(24):11704-11 PMID: 16357182
  39. An ATR- and Cdc7-dependent DNA damage checkpoint that inhibits initiation of DNA replication.
    Mol Cell. 2003 Jan;11(1):203-13 PMID: 12535533
  40. DNA interstrand cross-link repair in the cell cycle: a critical role for polymerase zeta in G1 phase.
    Cell Cycle. 2006 May;5(10):1044-7 PMID: 16687932
  41. DNA structure-induced recruitment and activation of the Fanconi anemia pathway protein FANCD2.
    Mol Cell Biol. 2007 Jun;27(12):4283-92 PMID: 17420278
  42. Identification of FAAP24, a Fanconi anemia core complex protein that interacts with FANCM.
    Mol Cell. 2007 Feb 9;25(3):331-43 PMID: 17289582
  43. Interstrand crosslink repair: can XPF-ERCC1 be let off the hook?
    Trends Genet. 2008 Feb;24(2):70-6 PMID: 18192062
  44. Fanconi anemia proteins are required to prevent accumulation of replication-associated DNA double-strand breaks.
    Mol Cell Biol. 2006 Jan;26(2):425-37 PMID: 16382135
  45. Role for DNA polymerase kappa in the processing of N2-N2-guanine interstrand cross-links.
    J Biol Chem. 2008 Jun 20;283(25):17075-82 PMID: 18434313
  46. The role of BRCA2 in replication-coupled DNA interstrand cross-link repair in vitro.
    Nat Struct Mol Biol. 2006 Aug;13(8):729-33 PMID: 16845393
  47. Distribution and repair of bipyrimidine photoproducts in solar UV-irradiated mammalian cells. Possible role of Dewar photoproducts in solar mutagenesis.
    J Biol Chem. 2000 Sep 1;275(35):26732-42 PMID: 10827179
  48. Interstrand cross-links induce DNA synthesis in damaged and undamaged plasmids in mammalian cell extracts.
    Mol Cell Biol. 1999 Aug;19(8):5619-30 PMID: 10409751
  49. Repair of an interstrand DNA cross-link initiated by ERCC1-XPF repair/recombination nuclease.
    J Biol Chem. 2000 Aug 25;275(34):26632-6 PMID: 10882712
  50. Fanconi anemia and DNA repair.
    Hum Mol Genet. 2001 Oct 1;10(20):2253-9 PMID: 11673408
  51. DNA replication is required To elicit cellular responses to psoralen-induced DNA interstrand cross-links.
    Mol Cell Biol. 2000 Nov;20(21):8283-9 PMID: 11027296
  52. Repair of DNA interstrand cross-links.
    Mutat Res. 2001 Sep 4;486(4):217-47 PMID: 11516927
  53. Distortion-dependent unhooking of interstrand cross-links in mammalian cell extracts.
    Biochemistry. 2008 Sep 16;47(37):9920-30 PMID: 18702509
Article Info
Journal
Molecular cell
Abbr.
Mol Cell
ISSN
1097-4164
DOI
10.1016/j.molcel.2009.08.014
Published
2009-09-11
Pages
704-15
Language
English
Region
United States
NLM ID
9802571
PMCID
PMC2756577
Subset
IM
Grants
NCI NIH HHS · R01 CA092245 · United States
NCI NIH HHS · R01 CA092245-08 · United States
NIGMS NIH HHS · R01 GM077495-02 · United States
PHS HHS · C020907 · United States
NIGMS NIH HHS · R01 GM077495 · United States
NIGMS NIH HHS · GM077495 · United States
NCI NIH HHS · CA92245 · United States
External Links
PubMed source DOI: 10.1016/j.molcel.2009.08.014 PMC Full Text View journal details
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