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PMID: 11904432 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

DNA-dependent protein kinase suppresses double-strand break-induced and spontaneous homologous recombination.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 99 ·No. 6 ·2002-03-19 ·Pages 3758-63

Allen C, Kurimasa A, Brenneman MA, Chen DJ, Nickoloff JA

Abstract

DNA-dependent protein kinase (DNA-PK), composed of Ku70, Ku80, and the catalytic subunit (DNA-PKcs), is involved in repairing double-strand breaks (DSBs) by nonhomologous end-joining (NHEJ). Certain proteins involved in NHEJ are also involved in DSB repair by homologous recombination (HR). To test the effects of DNA-PKcs on DSB-induced HR, we integrated neo direct repeat HR substrates carrying the I-SceI recognition sequence into DNA-PKcs-defective Chinese hamster ovary (V3) cells. The DNA-PKcs defect was complemented with a human DNA-PKcs cDNA. DSB-induced HR frequencies were 1.5- to 3-fold lower with DNA-PKcs complementation. In complemented and uncomplemented strains, all products arose by gene conversion without associated crossover, and average conversion tract lengths were similar. Suppression of DSB-induced HR in complemented cells probably reflects restoration of NHEJ, consistent with competition between HR and NHEJ during DSB repair. Interestingly, spontaneous HR rates were 1.6- to >3.5-fold lower with DNA-PKcs complementation. DNA-PKcs may suppress spontaneous HR through NHEJ of spontaneous DSBs, perhaps at stalled or blocked replication forks. Because replication protein A (RPA) is involved in both replication and HR, and is phosphorylated by DNA-PKcs, it is possible that the suppression of spontaneous HR by DNA-PKcs reflects regulation of replication-dependent HR by DNA-PKcs, perhaps by means of phosphorylation of RPA.

Keywords
Non-programmatic
MeSH Terms
Animals Blotting, Western Cell Line Cricetinae DNA/genetics,metabolism DNA Damage/genetics DNA Repair/genetics DNA Replication/genetics DNA-Activated Protein Kinase DNA-Binding Proteins Gene Conversion/genetics Genetic Complementation Test Humans Models, Genetic Mutation Neomycin Nuclear Proteins Protein Serine-Threonine Kinases/genetics,metabolism Recombination, Genetic/genetics Repetitive Sequences, Nucleic Acid/genetics Sequence Homology, Nucleic Acid
Chemicals
DNA-Binding Proteins Nuclear Proteins DNA DNA-Activated Protein Kinase PRKDC protein, human Protein Serine-Threonine Kinases Neomycin
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Allen Chris
Department of Molecular Genetics and Microbiology, University of New Mexico School of Medicine, Albuquerque, NM 87131, USA.
Kurimasa Akihiro
Brenneman Mark A
Chen David J
Nickoloff Jac A
References (72)
72 references, click to expand
  1. Relative rates of homologous and nonhomologous recombination in transfected DNA.
    Proc Natl Acad Sci U S A. 1985 May;82(10):3355-9 PMID: 2987922
  2. Heteroduplex deoxyribonucleic acid base mismatch repair in bacteria.
    Microbiol Rev. 1986 Jun;50(2):133-65 PMID: 3523187
  3. Nonhomologous recombination in mammalian cells: role for short sequence homologies in the joining reaction.
    Mol Cell Biol. 1986 Dec;6(12):4295-304 PMID: 3025650
  4. Differential effects of base-pair mismatch on intrachromosomal versus extrachromosomal recombination in mouse cells.
    Proc Natl Acad Sci U S A. 1987 Aug;84(15):5340-4 PMID: 3037544
  5. Double-strand breaks stimulate alternative mechanisms of recombination repair.
    J Mol Biol. 1989 Jun 5;207(3):527-41 PMID: 2668534
  6. The barrier to recombination between Escherichia coli and Salmonella typhimurium is disrupted in mismatch-repair mutants.
    Nature. 1989 Nov 23;342(6248):396-401 PMID: 2555716
  7. Homologous recombination in mammalian cells.
    Annu Rev Genet. 1989;23:199-225 PMID: 2694931
  8. Replication-mediated DNA damage by camptothecin induces phosphorylation of RPA by DNA-dependent protein kinase and dissociates RPA:DNA-PK complexes.
    EMBO J. 1999 Mar 1;18(5):1397-406 PMID: 10064605
  9. Increase of spontaneous intrachromosomal homologous recombination in mammalian cells expressing a mutant p53 protein.
    Oncogene. 1997 Mar 6;14(9):1117-22 PMID: 9070661
  10. Inactivation of p53 results in high rates of homologous recombination.
    Oncogene. 1997 Apr 17;14(15):1847-57 PMID: 9150391
  11. Stable DNA replication: interplay between DNA replication, homologous recombination, and transcription.
    Microbiol Mol Biol Rev. 1997 Jun;61(2):212-38 PMID: 9184011
  12. The DNA-dependent protein kinase catalytic activity regulates DNA end processing by means of Ku entry into DNA.
    J Biol Chem. 1999 Mar 19;274(12):7848-56 PMID: 10075677
  13. The role of the mismatch repair machinery in regulating mitotic and meiotic recombination between diverged sequences in yeast.
    Genetics. 1999 Apr;151(4):1299-313 PMID: 10101158
  14. Requirement for the kinase activity of human DNA-dependent protein kinase catalytic subunit in DNA strand break rejoining.
    Mol Cell Biol. 1999 May;19(5):3877-84 PMID: 10207111
  15. DNA-dependent protein kinase and related proteins.
    Biochem Soc Symp. 1999;64:91-104 PMID: 10207623
  16. Relocalization of telomeric Ku and SIR proteins in response to DNA strand breaks in yeast.
    Cell. 1999 May 28;97(5):621-33 PMID: 10367891
  17. 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
  18. Replication protein A (RPA): the eukaryotic SSB.
    Crit Rev Biochem Mol Biol. 1999;34(3):141-80 PMID: 10473346
  19. Mammalian XRCC2 promotes the repair of DNA double-strand breaks by homologous recombination.
    Nature. 1999 Sep 23;401(6751):397-9 PMID: 10517641
  20. A defect in mismatch repair in Saccharomyces cerevisiae stimulates ectopic recombination between homeologous genes by an excision repair dependent process.
    Genetics. 1990 Nov;126(3):535-47 PMID: 2249754
  21. Two alternative pathways of double-strand break repair that are kinetically separable and independently modulated.
    Mol Cell Biol. 1992 Mar;12(3):1292-303 PMID: 1545810
  22. Characterization of insertion mutations in the Saccharomyces cerevisiae MSH1 and MSH2 genes: evidence for separate mitochondrial and nuclear functions.
    Genetics. 1992 Dec;132(4):975-85 PMID: 1334021
  23. High spontaneous intrachromosomal recombination rates in ataxia-telangiectasia.
    Science. 1993 May 28;260(5112):1327-30 PMID: 8493577
  24. Gene conversions and crossing over during homologous and homeologous ectopic recombination in Saccharomyces cerevisiae.
    Genetics. 1993 Sep;135(1):5-16 PMID: 8224827
  25. Absence of p350 subunit of DNA-activated protein kinase from a radiosensitive human cell line.
    Science. 1995 Feb 24;267(5201):1183-5 PMID: 7855602
  26. Interspecies gene exchange in bacteria: the role of SOS and mismatch repair systems in evolution of species.
    Cell. 1995 Feb 10;80(3):507-15 PMID: 7859291
  27. Induction of homologous recombination in mammalian chromosomes by using the I-SceI system of Saccharomyces cerevisiae.
    Mol Cell Biol. 1995 Apr;15(4):1968-73 PMID: 7891691
  28. Loss of the catalytic subunit of the DNA-dependent protein kinase in DNA double-strand-break-repair mutant mammalian cells.
    Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3171-4 PMID: 7724535
  29. Mismatch correction acts as a barrier to homeologous recombination in Saccharomyces cerevisiae.
    Genetics. 1995 Mar;139(3):1175-88 PMID: 7768431
  30. Inactivation of the mouse Msh2 gene results in mismatch repair deficiency, methylation tolerance, hyperrecombination, and predisposition to cancer.
    Cell. 1995 Jul 28;82(2):321-30 PMID: 7628020
  31. DNA strand exchange mediated by a RAD51-ssDNA nucleoprotein filament with polarity opposite to that of RecA.
    Cell. 1995 Aug 11;82(3):453-61 PMID: 7634335
  32. scid cells are deficient in Ku and replication protein A phosphorylation by the DNA-dependent protein kinase.
    Mol Cell Biol. 1995 Oct;15(10):5700-6 PMID: 7565721
  33. Mitotic crossovers between diverged sequences are regulated by mismatch repair proteins in Saccaromyces cerevisiae.
    Mol Cell Biol. 1996 Mar;16(3):1085-93 PMID: 8622653
  34. Physical interaction between human RAD52 and RPA is required for homologous recombination in mammalian cells.
    J Biol Chem. 1996 Aug 2;271(31):18996-9000 PMID: 8702565
  35. Homologous and homeologous intermolecular gene conversion are not differentially affected by mutations in the DNA damage or the mismatch repair genes RAD1, RAD50, RAD51, RAD52, RAD54, PMS1 and MSH2.
    Genetics. 1996 Jun;143(2):755-67 PMID: 8725224
  36. Chromosomal double-strand break repair in Ku80-deficient cells.
    Proc Natl Acad Sci U S A. 1996 Aug 20;93(17):8929-33 PMID: 8799130
  37. The mismatch repair system reduces meiotic homeologous recombination and stimulates recombination-dependent chromosome loss.
    Mol Cell Biol. 1996 Nov;16(11):6110-20 PMID: 8887641
  38. The DNA damage response in DNA-dependent protein kinase-deficient SCID mouse cells: replication protein A hyperphosphorylation and p53 induction.
    Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):13825-30 PMID: 8943020
  39. A mutation in mouse rad51 results in an early embryonic lethal that is suppressed by a mutation in p53.
    Mol Cell Biol. 1996 Dec;16(12):7133-43 PMID: 8943369
  40. Influence of DNA sequence identity on efficiency of targeted gene replacement.
    Mol Cell Biol. 1997 Jan;17(1):278-86 PMID: 8972208
  41. DNA double-strand breaks caused by replication arrest.
    EMBO J. 1997 Jan 15;16(2):430-8 PMID: 9029161
  42. Fine-resolution analysis of products of intrachromosomal homeologous recombination in mammalian cells.
    Mol Cell Biol. 1997 Jul;17(7):3614-28 PMID: 9199296
  43. Dual roles for DNA sequence identity and the mismatch repair system in the regulation of mitotic crossing-over in yeast.
    Proc Natl Acad Sci U S A. 1997 Sep 2;94(18):9757-62 PMID: 9275197
  44. Chromosomal double-strand breaks induce gene conversion at high frequency in mammalian cells.
    Mol Cell Biol. 1997 Nov;17(11):6386-93 PMID: 9343400
  45. Interaction of p53 with the human Rad51 protein.
    Nucleic Acids Res. 1997 Oct 1;25(19):3868-74 PMID: 9380510
  46. Gene conversion tracts from double-strand break repair in mammalian cells.
    Mol Cell Biol. 1998 Jan;18(1):93-101 PMID: 9418857
  47. 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
  48. Studies of the interaction between Rad52 protein and the yeast single-stranded DNA binding protein RPA.
    Mol Cell Biol. 1998 Jul;18(7):4400-6 PMID: 9632824
  49. Rad52 associates with RPA and functions with rad55 and rad57 to assemble meiotic recombination complexes.
    Genes Dev. 1998 Jul 15;12(14):2208-21 PMID: 9679065
  50. Role of the human RAD51 protein in homologous recombination and double-stranded-break repair.
    Trends Biochem Sci. 1998 Jul;23(7):247-51 PMID: 9697414
  51. 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
  52. Homologous recombination, but not DNA repair, is reduced in vertebrate cells deficient in RAD52.
    Mol Cell Biol. 1998 Nov;18(11):6430-5 PMID: 9774659
  53. DNA end-joining: from yeast to man.
    Trends Biochem Sci. 1998 Oct;23(10):394-8 PMID: 9810228
  54. XRCC3 promotes homology-directed repair of DNA damage in mammalian cells.
    Genes Dev. 1999 Oct 15;13(20):2633-8 PMID: 10541549
  55. Brca1 controls homology-directed DNA repair.
    Mol Cell. 1999 Oct;4(4):511-8 PMID: 10549283
  56. Multiple pathways for repair of DNA double-strand breaks in mammalian chromosomes.
    Mol Cell Biol. 1999 Dec;19(12):8353-60 PMID: 10567560
  57. DNA damage-induced cell cycle checkpoints and DNA strand break repair in development and tumorigenesis.
    Oncogene. 1999 Dec 20;18(55):7883-99 PMID: 10630641
  58. Activation of DNA-dependent protein kinase by single-stranded DNA ends.
    J Biol Chem. 2000 Jan 21;275(3):1541-50 PMID: 10636842
  59. Replication fork pausing and recombination or "gimme a break".
    Genes Dev. 2000 Jan 1;14(1):1-10 PMID: 10640269
  60. Regulation of double-strand break-induced mammalian homologous recombination by UBL1, a RAD51-interacting protein.
    Nucleic Acids Res. 2000 Mar 1;28(5):1145-53 PMID: 10666456
  61. XRCC3 is required for efficient repair of chromosome breaks by homologous recombination.
    Mutat Res. 2000 Mar 20;459(2):89-97 PMID: 10725659
  62. Homologous and non-homologous recombination differentially affect DNA damage repair in mice.
    EMBO J. 2000 Apr 3;19(7):1703-10 PMID: 10747037
  63. Mouse RAD54 affects DNA double-strand break repair and sister chromatid exchange.
    Mol Cell Biol. 2000 May;20(9):3147-56 PMID: 10757799
  64. BASC, a super complex of BRCA1-associated proteins involved in the recognition and repair of aberrant DNA structures.
    Genes Dev. 2000 Apr 15;14(8):927-39 PMID: 10783165
  65. Partners and pathwaysrepairing a double-strand break.
    Trends Genet. 2000 Jun;16(6):259-64 PMID: 10827453
  66. The Rad51 paralog Rad51B promotes homologous recombinational repair.
    Mol Cell Biol. 2000 Sep;20(17):6476-82 PMID: 10938124
  67. Reconstitution of the strand invasion step of double-strand break repair using human Rad51 Rad52 and RPA proteins.
    J Mol Biol. 2000 Nov 24;304(2):151-64 PMID: 11080452
  68. Homologous recombinational repair of double-strand breaks in yeast is enhanced by MAT heterozygosity through yKU-dependent and -independent mechanisms.
    Genetics. 2001 Feb;157(2):579-89 PMID: 11156980
  69. Basis for avid homologous DNA strand exchange by human Rad51 and RPA.
    J Biol Chem. 2001 Mar 23;276(12):8798-806 PMID: 11124265
  70. Chromosome instability and defective recombinational repair in knockout mutants of the five Rad51 paralogs.
    Mol Cell Biol. 2001 Apr;21(8):2858-66 PMID: 11283264
  71. 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
  72. Model for homologous recombination during transfer of DNA into mouse L cells: role for DNA ends in the recombination process.
    Mol Cell Biol. 1984 Jun;4(6):1020-34 PMID: 6330525
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.052545899
Published
2002-03-19
Pages
3758-63
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC122597
Subset
IM
Grants
NCI NIH HHS · R01 CA077693 · United States
NCI NIH HHS · CA74046 · United States
NCI NIH HHS · CA77693 · United States
External Links
PubMed source DOI: 10.1073/pnas.052545899 PMC Full Text View journal details
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