title: Mammalian polymerase θ promotes alternative NHEJ and suppresses recombination.
abstract: The alternative non-homologous end-joining (NHEJ) machinery facilitates several genomic rearrangements, some of which can lead to cellular transformation. This error-prone repair pathway is triggered upon telomere de-protection to promote the formation of deleterious chromosome end-to-end fusions. Using next-generation sequencing technology, here we show that repair by alternative NHEJ yields non-TTAGGG nucleotide insertions at fusion breakpoints of dysfunctional telomeres. Investigating the enzymatic activity responsible for the random insertions enabled us to identify polymerase theta (Polθ; encoded by Polq in mice) as a crucial alternative NHEJ factor in mammalian cells. Polq inhibition suppresses alternative NHEJ at dysfunctional telomeres, and hinders chromosomal translocations at non-telomeric loci. In addition, we found that loss of Polq in mice results in increased rates of homology-directed repair, evident by recombination of dysfunctional telomeres and accumulation of RAD51 at double-stranded breaks. Lastly, we show that depletion of Polθ has a synergistic effect on cell survival in the absence of BRCA genes, suggesting that the inhibition of this mutagenic polymerase represents a valid therapeutic avenue for tumours carrying mutations in homology-directed repair genes.
authors: Skirball Institute of Biomolecular Medicine, Department of Cell Biology, NYU School of Medicine, New York, New York 10016, USA.
authors: Pedro A
authors: PA
authors: Mateos-Gomez
authors: Department of Molecular Biosciences, Institute for Cellular and Molecular Biology, University of Texas at Austin. 2506 Speedway Stop A5000, Austin, Texas 78712, USA.
authors: Fade
authors: F
authors: Gong
authors: Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California 92037, USA.
authors: Nidhi
authors: N
authors: Nair
authors: Department of Molecular Biosciences, Institute for Cellular and Molecular Biology, University of Texas at Austin. 2506 Speedway Stop A5000, Austin, Texas 78712, USA.
authors: Kyle M
authors: KM
authors: Miller
authors: Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California 92037, USA.
authors: Eros
authors: E
authors: Lazzerini-Denchi
authors: Skirball Institute of Biomolecular Medicine, Department of Cell Biology, NYU School of Medicine, New York, New York 10016, USA.
authors: Agnel
authors: A
authors: Sfeir
mesh_terms: D000818:Animals; D001483:Base Sequence; D016923:Cell Death / Q000235:genetics; D002460:Cell Line; D002869:Chromosome Aberrations; D033481:Chromosomes, Mammalian / Q000235:genetics / Q000378:metabolism*; D053903:DNA Breaks, Double-Stranded; D059766:DNA End-Joining Repair; D004259:DNA-Directed DNA Polymerase / Q000172:deficiency / Q000378:metabolism*; D019398:Genes, BRCA1; D024522:Genes, BRCA2; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases / Q000235:genetics / Q000378:metabolism; D051135:Rad51 Recombinase / Q000378:metabolism; D011995:Recombination, Genetic / Q000235:genetics; D059767:Recombinational DNA Repair / Q000235:genetics; D016615:Telomere / Q000235:genetics* / Q000378:metabolism*; D014178:Translocation, Genetic / Q000235:genetics; D000097462:DNA Polymerase theta
chemical_list: D004259:DNA-Directed DNA Polymerase; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases; D051135:Rad51 Recombinase; D000097462:DNA Polymerase theta; C499320:PARP1 protein, human
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