title: DNA ligase IV and artemis act cooperatively to suppress homologous recombination in human cells: implications for DNA double-strand break repair.
abstract: Nonhomologous end-joining (NHEJ) and homologous recombination (HR) are two major pathways for repairing DNA double-strand breaks (DSBs); however, their respective roles in human somatic cells remain to be elucidated. Here we show using a series of human gene-knockout cell lines that NHEJ repairs nearly all of the topoisomerase II- and low-dose radiation-induced DNA damage, while it negatively affects survival of cells harbouring replication-associated DSBs. Intriguingly, we find that loss of DNA ligase IV, a critical NHEJ ligase, and Artemis, an NHEJ factor with endonuclease activity, independently contribute to increased resistance to replication-associated DSBs. We also show that loss of Artemis alleviates hypersensitivity of DNA ligase IV-null cells to low-dose radiation- and topoisomerase II-induced DSBs. Finally, we demonstrate that Artemis-null human cells display increased gene-targeting efficiencies, particularly in the absence of DNA ligase IV. Collectively, these data suggest that DNA ligase IV and Artemis act cooperatively to promote NHEJ, thereby suppressing HR. Our results point to the possibility that HR can only operate on accidental DSBs when NHEJ is missing or abortive, and Artemis may be involved in pathway switching from incomplete NHEJ to HR.
authors: Graduate School of Nanobioscience, Yokohama City University, Yokohama, Japan.
authors: Aya
authors: A
authors: Kurosawa
authors: Shinta
authors: S
authors: Saito
authors: Sairei
authors: S
authors: So
authors: Mitsumasa
authors: M
authors: Hashimoto
authors: Kuniyoshi
authors: K
authors: Iwabuchi
authors: Haruka
authors: H
authors: Watabe
authors: Noritaka
authors: N
authors: Adachi
mesh_terms: D002460:Cell Line; D053903:DNA Breaks, Double-Stranded / Q000187:drug effects / Q000528:radiation effects; D059766:DNA End-Joining Repair / Q000187:drug effects / Q000528:radiation effects; D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000172:deficiency / Q000378:metabolism*; D004261:DNA Replication / Q000187:drug effects / Q000528:radiation effects; D004250:DNA Topoisomerases, Type II / Q000378:metabolism; D004268:DNA-Binding Proteins; D004720:Endonucleases; D055786:Gene Knockout Techniques; D018390:Gene Targeting; D059765:Homologous Recombination / Q000187:drug effects / Q000528:radiation effects; D006801:Humans; D009154:Mutation; D009687:Nuclear Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D059005:Topoisomerase II Inhibitors / Q000494:pharmacology
chemical_list: D004268:DNA-Binding Proteins; C000607893:LIG4 protein, human; D009687:Nuclear Proteins; D059005:Topoisomerase II Inhibitors; C426344:DCLRE1C protein, human; D004720:Endonucleases; D004250:DNA Topoisomerases, Type II; D011088:DNA Ligases; D000072481:DNA Ligase ATP
references: Biochem Biophys Res Commun. 2008 Aug 8;372(4):662-7
references: 18538133
references: J Biol Chem. 2004 Sep 3;279(36):37343-8
references: 15218034
references: Front Med. 2012 Jun;6(2):156-64
references: 22660976
references: DNA Cell Biol. 2006 Jan;25(1):19-24
references: 16405397
references: J Biol Chem. 2003 Sep 19;278(38):35897-902
references: 12842886
references: Cell. 2002 Mar 22;108(6):781-94
references: 11955432
references: EMBO J. 1999 Mar 1;18(5):1397-406
references: 10064605
references: Mol Cell Biol. 2007 Apr;27(7):2562-71
references: 17242200
references: Nature. 2007 Sep 27;449(7161):483-6
references: 17898768
references: Annu Rev Biochem. 2010;79:181-211
references: 20192759
references: Nat Rev Genet. 2006 Jan;7(1):45-54
references: 16369571
references: Annu Rev Genet. 2011;45:247-71
references: 21910633
references: Cancer Sci. 2005 Feb;96(2):134-41
references: 15723659
references: EMBO J. 2011 Mar 16;30(6):1079-92
references: 21317870
references: Nucleic Acids Res. 2008 Nov;36(19):6333-42
references: 18835848
references: Mol Cell. 1998 Oct;2(4):477-84
references: 9809069
references: Genes Dev. 2004 Jun 1;18(11):1283-92
references: 15175260
references: Radiother Oncol. 2011 Oct;101(1):7-12
references: 21737170
references: Chem Rev. 2009 Jul;109(7):2894-902
references: 19476377
references: Nature. 2005 Apr 14;434(7035):917-21
references: 15829967
references: DNA Repair (Amst). 2005 Jul 12;4(7):749-59
references: 15978533
references: Mol Cell. 2012 Aug 24;47(4):497-510
references: 22920291
references: J Biol Chem. 2003 May 30;278(22):20303-12
references: 12660252
references: Biochim Biophys Acta. 2011 Sep;1809(9):509-23
references: 21704205
references: Proc Natl Acad Sci U S A. 2001 Mar 13;98(6):3243-8
references: 11248063
references: Nat Rev Cancer. 2009 May;9(5):338-50
references: 19377506
references: Nat Cell Biol. 2007 Jun;9(6):675-82
references: 17486118
references: Proc Natl Acad Sci U S A. 2008 Jun 24;105(25):8703-8
references: 18562296
references: J Biol Chem. 2007 Apr 6;282(14):10138-45
references: 17272272
references: Curr Opin Genet Dev. 2007 Oct;17(5):435-42
references: 17905579
references: J Biol Chem. 2008 Aug 29;283(35):23711-20
references: 18596031
references: FEBS Lett. 2010 Sep 10;584(17):3703-8
references: 20691183
references: J Biol Chem. 2007 Mar 2;282(9):6582-7
references: 17189255
references: EMBO J. 1998 Sep 15;17(18):5497-508
references: 9736627
references: DNA Repair (Amst). 2006 Mar 7;5(3):303-11
references: 16325483
references: Nature. 2007 Sep 27;449(7161):478-82
references: 17713479
references: Proc Natl Acad Sci U S A. 2002 Jan 22;99(2):832-7
references: 11792868
references: DNA Repair (Amst). 2010 Dec 10;9(12):1307-14
references: 21030321
references: Mol Cell Biol. 2005 Dec;25(24):10842-52
references: 16314509
references: EMBO Rep. 2008 Jan;9(1):91-6
references: 18064046
references: Nature. 1999 Apr 22;398(6729):728-31
references: 10227297
references: Nucleic Acids Res. 2012 Jul;40(13):5795-818
references: 22467216
references: Biotechniques. 2006 Sep;41(3):311-6
references: 16989091
references: EMBO J. 2004 May 5;23(9):1987-97
references: 15071507
references: EMBO J. 2006 Mar 22;25(6):1305-14
references: 16498404
references: Genes Cells. 2006 Aug;11(8):935-48
references: 16866876
references: DNA Repair (Amst). 2012 Oct 1;11(10):781-8
references: 22889934
references: DNA Repair (Amst). 2002 Dec 5;1(12):1017-26
references: 12531011
references: J Radiat Res. 2010;51(5):503-9
references: 20543526
references: Annu Rev Biochem. 1989;58:351-75
references: 2549853
references: Mol Cell Biol. 1998 Nov;18(11):6430-5
references: 9774659
references: EMBO Rep. 2005 Aug;6(8):729-35
references: 16025133
references: PLoS Genet. 2011 Jun;7(6):e1002080
references: 21655080
references: DNA Repair (Amst). 2005 Jul 12;4(7):845-51
references: 15936993
references: Mol Cell. 2004 Dec 3;16(5):715-24
references: 15574327
references: Proc Natl Acad Sci U S A. 2001 Oct 9;98(21):12109-13
references: 11593023
references: Mol Cell Biol. 2004 Oct;24(20):9207-20
references: 15456891
references: Nucleic Acids Res. 2003 Aug 1;31(15):4373-84
references: 12888496
references: DNA Repair (Amst). 2006 Sep 8;5(9-10):1199-212
references: 16822725
references: Cell. 2006 Jan 27;124(2):260-2
references: 16439201
references: DNA Cell Biol. 2008 Jan;27(1):55-61
references: 17941805
references: Cell. 2010 Apr 16;141(2):243-54
references: 20362325