title: Chromosomal translocations in human cells are generated by canonical nonhomologous end-joining.
abstract: Breakpoint junctions of the chromosomal translocations that occur in human cancers display hallmarks of nonhomologous end-joining (NHEJ). In mouse cells, translocations are suppressed by canonical NHEJ (c-NHEJ) components, which include DNA ligase IV (LIG4), and instead arise from alternative NHEJ (alt-NHEJ). Here we used designer nucleases (ZFNs, TALENs, and CRISPR/Cas9) to introduce DSBs on two chromosomes to study translocation joining mechanisms in human cells. Remarkably, translocations were altered in cells deficient for LIG4 or its interacting protein XRCC4. Translocation junctions had significantly longer deletions and more microhomology, indicative of alt-NHEJ. Thus, unlike mouse cells, translocations in human cells are generated by c-NHEJ. Human cancer translocations induced by paired Cas9 nicks also showed a dependence on c-NHEJ, despite having distinct joining characteristics. These results demonstrate an unexpected and striking species-specific difference for common genomic rearrangements associated with tumorigenesis.
authors.affiliation: Museum National d'Histoire Naturelle, 43 rue Cuvier, F-75005 Paris, France; CNRS, UMR7196, 43 rue Cuvier, F-75005 Paris, France; Inserm, U1154, 43 rue Cuvier, F-75005 Paris, France.
authors.forename: Hind
authors.initials: H
authors.lastname: Ghezraoui
authors.affiliation: Museum National d'Histoire Naturelle, 43 rue Cuvier, F-75005 Paris, France; CNRS, UMR7196, 43 rue Cuvier, F-75005 Paris, France; Inserm, U1154, 43 rue Cuvier, F-75005 Paris, France.
authors.forename: Marion
authors.initials: M
authors.lastname: Piganeau
authors.affiliation: Museum National d'Histoire Naturelle, 43 rue Cuvier, F-75005 Paris, France; CNRS, UMR7196, 43 rue Cuvier, F-75005 Paris, France; Inserm, U1154, 43 rue Cuvier, F-75005 Paris, France.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Renouf
authors.affiliation: Museum National d'Histoire Naturelle, 43 rue Cuvier, F-75005 Paris, France; CNRS, UMR7196, 43 rue Cuvier, F-75005 Paris, France; Inserm, U1154, 43 rue Cuvier, F-75005 Paris, France.
authors.forename: Jean-Baptiste
authors.initials: JB
authors.lastname: Renaud
authors.affiliation: Department of Internal Medicine and University of New Mexico Cancer Center, University of New Mexico, Albuquerque, NM 87131, USA.
authors.forename: Annahita
authors.initials: A
authors.lastname: Sallmyr
authors.affiliation: Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota Medical School, Minneapolis, MN 55455, USA.
authors.forename: Brian
authors.initials: B
authors.lastname: Ruis
authors.affiliation: Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota Medical School, Minneapolis, MN 55455, USA.
authors.forename: Sehyun
authors.initials: S
authors.lastname: Oh
authors.affiliation: Department of Internal Medicine and University of New Mexico Cancer Center, University of New Mexico, Albuquerque, NM 87131, USA.
authors.forename: Alan E
authors.initials: AE
authors.lastname: Tomkinson
authors.affiliation: Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota Medical School, Minneapolis, MN 55455, USA.
authors.forename: Eric A
authors.initials: EA
authors.lastname: Hendrickson
authors.affiliation: Museum National d'Histoire Naturelle, 43 rue Cuvier, F-75005 Paris, France; CNRS, UMR7196, 43 rue Cuvier, F-75005 Paris, France; Inserm, U1154, 43 rue Cuvier, F-75005 Paris, France.
authors.forename: Carine
authors.initials: C
authors.lastname: Giovannangeli
authors.affiliation: Developmental Biology Program, Memorial Sloan-Kettering Cancer Center, New York, NY 10065, USA. Electronic address: m-jasin@ski.mskcc.org.
authors.forename: Maria
authors.initials: M
authors.lastname: Jasin
authors.affiliation: Museum National d'Histoire Naturelle, 43 rue Cuvier, F-75005 Paris, France; CNRS, UMR7196, 43 rue Cuvier, F-75005 Paris, France; Inserm, U1154, 43 rue Cuvier, F-75005 Paris, France. Electronic address: ebrunet@mnhn.fr.
authors.forename: Erika
authors.initials: E
authors.lastname: Brunet
mesh_terms: D000818:Animals; D002877:Chromosomes, Human; D059766:DNA End-Joining Repair; D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics*; D003851:Deoxyribonucleases / Q000502:physiology*; D006801:Humans; D051379:Mice; D017384:Sequence Deletion; D013045:Species Specificity; D014178:Translocation, Genetic / Q000235:genetics*; D014407:Tumor Cells, Cultured
chemical_list: D004268:DNA-Binding Proteins; C000607893:LIG4 protein, human; C101467:XRCC4 protein, human; D003851:Deoxyribonucleases; D011088:DNA Ligases; D000072481:DNA Ligase ATP