Home LiteratureArticle Details
PMID: 15175261 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Collaboration of homologous recombination and nonhomologous end-joining factors for the survival and integrity of mice and cells.

Genes & development ·Vol. 18 ·No. 11 ·2004-06-01 ·Pages 1293-304

Couëdel C, Mills KD, Barchi M, Shen L, Olshen A, Johnson RD, Nussenzweig A, Essers J, Kanaar R, Li GC, Alt FW, Jasin M

Abstract

Homologous recombination (HR) and nonhomologous end-joining (NHEJ) are mechanistically distinct DNA repair pathways that contribute substantially to double-strand break (DSB) repair in mammalian cells. We have combined mutations in factors from both repair pathways, the HR protein Rad54 and the DNA-end-binding factor Ku80, which has a role in NHEJ. Rad54(-/-)Ku80(-/-) mice were severely compromised in their survival, such that fewer double mutants were born than expected, and only a small proportion of those born reached adulthood. However, double-mutant mice died at lower frequency from tumors than Ku80 single mutant mice, likely as a result of rapid demise at a young age from other causes. When challenged with an exogenous DNA damaging agent, ionizing radiation, double-mutant mice were exquisitely sensitive to low doses. Tissues and cells from double-mutant mice also showed indications of spontaneous DNA damage. Testes from some Rad54(-/-)Ku80(-/-) mice displayed enhanced apoptosis and reduced sperm production, and embryonic fibroblasts from Rad54(-/-)Ku80(-/-) animals accumulated foci of gamma-H2AX, a marker for DSBs. The substantially increased DNA damage response in the double mutants implies a cooperation of the two DSB repair pathways for survival and genomic integrity in the animal.

MeSH Terms
Aging/genetics Animals Animals, Newborn Antigens, Nuclear/genetics,metabolism Brain/pathology Cells, Cultured DNA Damage/genetics DNA Helicases DNA Repair/physiology DNA-Binding Proteins/genetics,metabolism Female Fetal Death/genetics Fibroblasts/physiology,radiation effects Histones/genetics,metabolism Ku Autoantigen Male Mice Mice, Mutant Strains Nuclear Proteins/genetics,metabolism Radiation, Ionizing Recombination, Genetic Survival Rate Testis/pathology
Chemicals
Antigens, Nuclear DNA-Binding Proteins Histones Nuclear Proteins gamma-H2AX protein, mouse DNA Helicases Rad54l protein, mouse Xrcc6 protein, mouse Ku Autoantigen
Authors & Affiliations
12 authors, click to expand affiliations / ORCID
Couëdel Chrystelle
Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, New York, New York 10021, USA.
Mills Kevin D
Barchi Marco
Shen Lingbo
Olshen Adam
Johnson Roger D
Nussenzweig André
Essers Jeroen
Kanaar Roland
Li Gloria C
Alt Frederick W
Jasin Maria
References (42)
42 references, click to expand
  1. Genetic analysis of the DNA-dependent protein kinase reveals an inhibitory role of Ku in late S-G2 phase DNA double-strand break repair.
    J Biol Chem. 2001 Nov 30;276(48):44413-8 PMID: 11577093
  2. Double-strand breaks and tumorigenesis.
    Trends Cell Biol. 2001 Nov;11(11):S52-9 PMID: 11684443
  3. Recombination at double-strand breaks and DNA ends: conserved mechanisms from phage to humans.
    Mol Cell. 2001 Dec;8(6):1163-74 PMID: 11779493
  4. DNA-dependent protein kinase suppresses double-strand break-induced and spontaneous homologous recombination.
    Proc Natl Acad Sci U S A. 2002 Mar 19;99(6):3758-63 PMID: 11904432
  5. Mammalian Ku86 mediates chromosomal fusions and apoptosis caused by critically short telomeres.
    EMBO J. 2002 May 1;21(9):2207-19 PMID: 11980718
  6. Increased ionizing radiation sensitivity and genomic instability in the absence of histone H2AX.
    Proc Natl Acad Sci U S A. 2002 Jun 11;99(12):8173-8 PMID: 12034884
  7. An xrcc4 defect or Wortmannin stimulates homologous recombination specifically induced by double-strand breaks in mammalian cells.
    Nucleic Acids Res. 2002 Aug 1;30(15):3454-63 PMID: 12140331
  8. Evidence for replicative repair of DNA double-strand breaks leading to oncogenic translocation and gene amplification.
    J Exp Med. 2002 Aug 19;196(4):469-80 PMID: 12186839
  9. Role of RAD52 epistasis group genes in homologous recombination and double-strand break repair.
    Microbiol Mol Biol Rev. 2002 Dec;66(4):630-70, table of contents PMID: 12456786
  10. Homologous repair of DNA damage and tumorigenesis: the BRCA connection.
    Oncogene. 2002 Dec 16;21(58):8981-93 PMID: 12483514
  11. Overexpression of Cu/Zn superoxide dismutase is lethal for mice lacking double-strand break repair.
    DNA Repair (Amst). 2003 Mar 1;2(3):285-94 PMID: 12547391
  12. Molecular views of recombination proteins and their control.
    Nat Rev Mol Cell Biol. 2003 Jun;4(6):435-45 PMID: 12778123
  13. The role of DNA breaks in genomic instability and tumorigenesis.
    Immunol Rev. 2003 Aug;194:77-95 PMID: 12846809
  14. Pathways of DNA double-strand break repair during the mammalian cell cycle.
    Mol Cell Biol. 2003 Aug;23(16):5706-15 PMID: 12897142
  15. Rapid, stabilizing palindrome rearrangements in somatic cells by the center-break mechanism.
    Mol Cell Biol. 2003 Dec;23(23):8740-50 PMID: 14612414
  16. Rad54 and DNA Ligase IV cooperate to maintain mammalian chromatid stability.
    Genes Dev. 2004 Jun 1;18(11):1283-92 PMID: 15175260
  17. Introduction of double-strand breaks into the genome of mouse cells by expression of a rare-cutting endonuclease.
    Mol Cell Biol. 1994 Dec;14(12):8096-106 PMID: 7969147
  18. Requirement for Ku80 in growth and immunoglobulin V(D)J recombination.
    Nature. 1996 Aug 8;382(6591):551-5 PMID: 8700231
  19. Disruption of mouse RAD54 reduces ionizing radiation resistance and homologous recombination.
    Cell. 1997 Apr 18;89(2):195-204 PMID: 9108475
  20. Growth retardation and leaky SCID phenotype of Ku70-deficient mice.
    Immunity. 1997 Nov;7(5):653-65 PMID: 9390689
  21. Hypersensitivity of Ku80-deficient cell lines and mice to DNA damage: the effects of ionizing radiation on growth, survival, and development.
    Proc Natl Acad Sci U S A. 1997 Dec 9;94(25):13588-93 PMID: 9391070
  22. DNA double-stranded breaks induce histone H2AX phosphorylation on serine 139.
    J Biol Chem. 1998 Mar 6;273(10):5858-68 PMID: 9488723
  23. Homology-directed repair is a major double-strand break repair pathway in mammalian cells.
    Proc Natl Acad Sci U S A. 1998 Apr 28;95(9):5172-7 PMID: 9560248
  24. Ku70: a candidate tumor suppressor gene for murine T cell lymphoma.
    Mol Cell. 1998 Jul;2(1):1-8 PMID: 9702186
  25. Homologous recombination and non-homologous end-joining pathways of DNA double-strand break repair have overlapping roles in the maintenance of chromosomal integrity in vertebrate cells.
    EMBO J. 1998 Sep 15;17(18):5497-508 PMID: 9736627
  26. Multiple pathways of recombination induced by double-strand breaks in Saccharomyces cerevisiae.
    Microbiol Mol Biol Rev. 1999 Jun;63(2):349-404 PMID: 10357855
  27. The Drosophila melanogaster DmRAD54 gene plays a crucial role in double-strand break repair after P-element excision and acts synergistically with Ku70 in the repair of X-ray damage.
    Mol Cell Biol. 1999 Sep;19(9):6269-75 PMID: 10454573
  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. Defective embryonic neurogenesis in Ku-deficient but not DNA-dependent protein kinase catalytic subunit-deficient mice.
    Proc Natl Acad Sci U S A. 2000 Mar 14;97(6):2668-73 PMID: 10716994
  30. Homologous and non-homologous recombination differentially affect DNA damage repair in mice.
    EMBO J. 2000 Apr 3;19(7):1703-10 PMID: 10747037
  31. Mouse RAD54 affects DNA double-strand break repair and sister chromatid exchange.
    Mol Cell Biol. 2000 May;20(9):3147-56 PMID: 10757799
  32. DNA repair protein Ku80 suppresses chromosomal aberrations and malignant transformation.
    Nature. 2000 Mar 30;404(6777):510-4 PMID: 10761921
  33. Sister chromatid gene conversion is a prominent double-strand break repair pathway in mammalian cells.
    EMBO J. 2000 Jul 3;19(13):3398-407 PMID: 10880452
  34. Coupled homologous and nonhomologous repair of a double-strand break preserves genomic integrity in mammalian cells.
    Mol Cell Biol. 2000 Dec;20(23):9068-75 PMID: 11074004
  35. Xrcc2 is required for genetic stability, embryonic neurogenesis and viability in mice.
    EMBO J. 2000 Dec 15;19(24):6675-85 PMID: 11118202
  36. Recombinational DNA double-strand breaks in mice precede synapsis.
    Nat Genet. 2001 Mar;27(3):271-6 PMID: 11242108
  37. Regulation of Saccharomyces Rad53 checkpoint kinase during adaptation from DNA damage-induced G2/M arrest.
    Mol Cell. 2001 Feb;7(2):293-300 PMID: 11239458
  38. Chromosomal stability and the DNA double-stranded break connection.
    Nat Rev Genet. 2001 Mar;2(3):196-206 PMID: 11256071
  39. Characterization of homologous recombination induced by replication inhibition in mammalian cells.
    EMBO J. 2001 Jul 16;20(14):3861-70 PMID: 11447127
  40. Repeat expansion by homologous recombination in the mouse germ line at palindromic sequences.
    Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8326-33 PMID: 11459971
  41. Mechanism and control of meiotic recombination initiation.
    Curr Top Dev Biol. 2001;52:1-53 PMID: 11529427
  42. Ku DNA end-binding protein modulates homologous repair of double-strand breaks in mammalian cells.
    Genes Dev. 2001 Dec 15;15(24):3237-42 PMID: 11751629
Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.1209204
Published
2004-06-01
Pages
1293-304
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC420355
Subset
IM
Grants
NCI NIH HHS · CA78497 · United States
NICHD NIH HHS · R01 HD040916 · United States
NIGMS NIH HHS · GM54668 · United States
NCI NIH HHS · CA56909 · United States
NCI NIH HHS · R01 CA056909 · United States
NICHD NIH HHS · HD40916 · United States
NIGMS NIH HHS · R01 GM054668 · United States
External Links
PubMed source DOI: 10.1101/gad.1209204 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2