title: RAG2 and XLF/Cernunnos interplay reveals a novel role for the RAG complex in DNA repair.
abstract: XRCC4-like factor (XLF) functions in classical non-homologous end-joining (cNHEJ) but is dispensable for the repair of DNA double-strand breaks (DSBs) generated during V(D)J recombination. A long-standing hypothesis proposes that, in addition to its canonical nuclease activity, the RAG1/2 proteins participate in the DNA repair phase of V(D)J recombination. Here we show that in the context of RAG2 lacking the C-terminus domain (Rag2(c/c) mice), XLF deficiency leads to a profound lymphopenia associated with a severe defect in V(D)J recombination and, in the absence of p53, increased genomic instability at V(D)J sites. In addition, Rag2(c/c) XLF(-/-) p53(-/-) mice develop aggressive pro-B cell lymphomas bearing complex chromosomal translocations and gene amplifications involving Igh and c-myc/pvt1 loci. Our results reveal an unanticipated functional interplay between the RAG complex and XLF in repairing RAG-induced DSBs and maintaining genome integrity during antigen receptor gene assembly.
authors.affiliation: Departments of Immunology and Genomes and Genetics, Institut Pasteur, CNRS-URA 1961, Paris 75015, France.
authors.forename: Chloé
authors.initials: C
authors.lastname: Lescale
authors.affiliation: Laboratory of Genome Dynamics in the Immune System, INSERM UMR1163, Université Paris Descartes Sorbonne Paris Cité, Institut Imagine, Paris 75015, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Abramowski
authors.affiliation: Departments of Immunology and Genomes and Genetics, Institut Pasteur, CNRS-URA 1961, Paris 75015, France.
authors.forename: Marie
authors.initials: M
authors.lastname: Bedora-Faure
authors.affiliation: Departments of Immunology and Genomes and Genetics, Institut Pasteur, CNRS-URA 1961, Paris 75015, France.
authors.forename: Valentine
authors.initials: V
authors.lastname: Murigneux
authors.affiliation: Laboratory of Genome Dynamics in the Immune System, INSERM UMR1163, Université Paris Descartes Sorbonne Paris Cité, Institut Imagine, Paris 75015, France.
authors.forename: Gabriella
authors.initials: G
authors.lastname: Vera
authors.affiliation: Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
authors.forename: David B
authors.initials: DB
authors.lastname: Roth
authors.affiliation: Laboratory of Genome Dynamics in the Immune System, INSERM UMR1163, Université Paris Descartes Sorbonne Paris Cité, Institut Imagine, Paris 75015, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Revy
authors.affiliation: Laboratory of Genome Dynamics in the Immune System, INSERM UMR1163, Université Paris Descartes Sorbonne Paris Cité, Institut Imagine, Paris 75015, France.
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: de Villartay
authors.affiliation: Departments of Immunology and Genomes and Genetics, Institut Pasteur, CNRS-URA 1961, Paris 75015, France.
authors.forename: Ludovic
authors.initials: L
authors.lastname: Deriano
mesh_terms: D000818:Animals; D053960:DNA Breaks; D004260:DNA Repair / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology*; D042822:Genomic Instability; D008214:Lymphocytes / Q000166:cytology / Q000502:physiology; D008231:Lymphopenia / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout
chemical_list: D004268:DNA-Binding Proteins; C499308:Rag2 protein, mouse; C519508:XLF protein, mouse
references.citation: Cell. 2004 Apr 16;117(2):171-84
references.pmid: 15084256
references.citation: Mol Cell. 2008 Sep 5;31(5):631-40
references.pmid: 18775323
references.citation: Annu Rev Genet. 2011;45:167-202
references.pmid: 21854230
references.citation: Proc Natl Acad Sci U S A. 1975 May;72(5):1932-6
references.pmid: 168584
references.citation: J Exp Med. 2010 Jul 5;207(7):1369-80
references.pmid: 20566716
references.citation: Mol Cell. 2001 Jan;7(1):77-87
references.pmid: 11172713
references.citation: Bioinformatics. 2012 Feb 1;28(3):423-5
references.pmid: 22155870
references.citation: J Biol Chem. 2006 May 19;281(20):13857-60
references.pmid: 16571728
references.citation: Nucleic Acids Res. 2004;32(15):4539-49
references.pmid: 15328366
references.citation: Nat Immunol. 2003 Jan;4(1):31-7
references.pmid: 12469118
references.citation: Microbiol Spectr. 2014 Dec;2(6). doi: 10.1128/microbiolspec.MDNA3-0041-2014
references.pmid: 26104458
references.citation: J Biol Chem. 2011 Sep 16;286(37):32638-50
references.pmid: 21775435
references.citation: J Vis Exp. 2013;(72):e50087
references.pmid: 23407477
references.citation: Nature. 1996 Aug 8;382(6591):551-5
references.pmid: 8700231
references.citation: Immunol Rev. 2004 Aug;200:115-31
references.pmid: 15242400
references.citation: Cell. 2002 Apr;109 Suppl:S45-55
references.pmid: 11983152
references.citation: Genome Res. 2009 Sep;19(9):1639-45
references.pmid: 19541911
references.citation: Nat Rev Mol Cell Biol. 2014 Jan;15(1):7-18
references.pmid: 24326623
references.citation: Immunity. 2002 Nov;17(5):639-51
references.pmid: 12433370
references.citation: J Immunol. 2011 Aug 15;187(4):1826-34
references.pmid: 21742970
references.citation: Curr Biol. 1994 Jan 1;4(1):1-7
references.pmid: 7922305
references.citation: Blood. 2011 Apr 14;117(15):4065-75
references.pmid: 21300982
references.citation: Genes Dev. 2002 Aug 1;16(15):1934-49
references.pmid: 12154124
references.citation: Mol Cell Biol. 2013 Sep;33(18):3568-79
references.pmid: 23836881
references.citation: Proc Natl Acad Sci U S A. 2007 May 8;104(19):7851-6
references.pmid: 17470781
references.citation: Cell. 2006 Jan 27;124(2):301-13
references.pmid: 16439205
references.citation: Annu Rev Immunol. 2012;30:175-202
references.pmid: 22224778
references.citation: Nucleic Acids Res. 2008 Oct;36(18):5750-62
references.pmid: 18776220
references.citation: Mol Cell. 2001 Jan;7(1):65-75
references.pmid: 11172712
references.citation: Cell Rep. 2013 Sep 12;4(5):870-8
references.pmid: 23994475
references.citation: Arch Immunol Ther Exp (Warsz). 2009 Mar-Apr;57(2):105-16
references.pmid: 19333736
references.citation: Nature. 2011 Mar 3;471(7336):119-23
references.pmid: 21368836
references.citation: Adv Immunol. 2005;86:43-112
references.pmid: 15705419
references.citation: Cell. 1992 Mar 6;68(5):855-67
references.pmid: 1547487
references.citation: Nat Immunol. 2007 Aug;8(8):801-8
references.pmid: 17641661
references.citation: Mol Cell. 2002 Dec;10(6):1379-90
references.pmid: 12504013
references.citation: Bioinformatics. 2012 Sep 15;28(18):i333-i339
references.pmid: 22962449
references.citation: Science. 2000 Dec 8;290(5498):1962-5
references.pmid: 11110662
references.citation: Nat Rev Immunol. 2003 Aug;3(8):656-66
references.pmid: 12974480
references.citation: Nucleic Acids Res. 2007;35(17):5755-62
references.pmid: 17717001
references.citation: Nucleic Acids Res. 2012 Feb;40(4):1684-94
references.pmid: 22228831
references.citation: Mol Cell. 2007 Dec 28;28(6):1093-101
references.pmid: 18158905
references.citation: Nature. 2007 Sep 27;449(7161):483-6
references.pmid: 17898768
references.citation: Cell. 2006 Jan 27;124(2):287-99
references.pmid: 16439204
references.citation: Mol Cell. 2001 Dec;8(6):1383-90
references.pmid: 11779512
references.citation: Cell. 2002 Jun 28;109(7):811-21
references.pmid: 12110179
references.citation: Mol Cell Biol. 2002 May;22(10):3460-73
references.pmid: 11971977
references.citation: Proc Natl Acad Sci U S A. 2007 Mar 13;104(11):4518-23
references.pmid: 17360556
references.citation: Proc Natl Acad Sci U S A. 2003 Feb 4;100(3):1209-14
references.pmid: 12531919
references.citation: Proc Natl Acad Sci U S A. 2011 Aug 2;108(31):12663-8
references.pmid: 21768349
references.citation: Nature. 2011 Jan 13;469(7329):250-4
references.pmid: 21160472
references.citation: Nature. 2011 Jan 13;469(7329):245-9
references.pmid: 21160476
references.citation: DNA Repair (Amst). 2014 Apr;16:11-22
references.pmid: 24674624
references.citation: Mol Cell Biol. 2013 Feb;33(4):701-11
references.pmid: 23207905
references.citation: Annu Rev Genet. 2013;47:433-55
references.pmid: 24050180
references.citation: Science. 2007 May 25;316(5828):1160-6
references.pmid: 17525332
references.citation: Immunity. 1997 Nov;7(5):653-65
references.pmid: 9390689
references.citation: Nucleic Acids Res. 2009 Feb;37(2):482-92
references.pmid: 19056826
references.citation: Bioinformatics. 2014 Apr 15;30(8):1064-1072
references.pmid: 24389656
references.citation: Bioinformatics. 2010 Aug 1;26(15):1895-6
references.pmid: 20639544
references.citation: Mol Cell Biol. 2014 Aug;34(15):2811-21
references.pmid: 24842905
references.citation: EMBO J. 2008 Jan 9;27(1):290-300
references.pmid: 18046455
references.citation: Adv Immunol. 2005;87:237-95
references.pmid: 16102576
references.citation: Bioinformatics. 2009 Jul 15;25(14):1754-60
references.pmid: 19451168
references.citation: Proc Natl Acad Sci U S A. 2015 May 19;112(20):E2575-84
references.pmid: 25941401
references.citation: Cell. 1996 Jul 12;86(1):159-71
references.pmid: 8689683
references.citation: Science. 1994 Sep 2;265(5177):1442-5
references.pmid: 8073286
references.citation: Nature. 2006 Jul 27;442(7101):466-70
references.pmid: 16799570
references.citation: Nucleic Acids Res. 2014 Jun;42(10):6352-64
references.pmid: 24753404