ATM orchestrates the DNA-damage response to counter toxic non-homologous end-joining at broken replication forks

Nat Commun. 2019 Jan 8;10(1):87. doi: 10.1038/s41467-018-07729-2.

Abstract

Mutations in the ATM tumor suppressor gene confer hypersensitivity to DNA-damaging chemotherapeutic agents. To explore genetic resistance mechanisms, we performed genome-wide CRISPR-Cas9 screens in cells treated with the DNA topoisomerase I inhibitor topotecan. Thus, we here establish that inactivating terminal components of the non-homologous end-joining (NHEJ) machinery or of the BRCA1-A complex specifically confer topotecan resistance to ATM-deficient cells. We show that hypersensitivity of ATM-mutant cells to topotecan or the poly-(ADP-ribose) polymerase (PARP) inhibitor olaparib reflects delayed engagement of homologous recombination at DNA-replication-fork associated single-ended double-strand breaks (DSBs), allowing some to be subject to toxic NHEJ. Preventing DSB ligation by NHEJ, or enhancing homologous recombination by BRCA1-A complex disruption, suppresses this toxicity, highlighting a crucial role for ATM in preventing toxic LIG4-mediated chromosome fusions. Notably, suppressor mutations in ATM-mutant backgrounds are different to those in BRCA1-mutant scenarios, suggesting new opportunities for patient stratification and additional therapeutic vulnerabilities for clinical exploitation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology*
  • Antineoplastic Agents / therapeutic use
  • Ataxia Telangiectasia Mutated Proteins / genetics*
  • Ataxia Telangiectasia Mutated Proteins / metabolism
  • BRCA1 Protein / metabolism
  • CRISPR-Cas Systems / genetics
  • Cell Line, Tumor
  • Cell Survival / genetics
  • DNA Breaks, Double-Stranded / drug effects
  • DNA End-Joining Repair / genetics*
  • DNA Ligase ATP / metabolism
  • DNA Replication / drug effects
  • DNA Replication / genetics
  • Drug Resistance, Neoplasm / genetics*
  • Female
  • Humans
  • Mice
  • Mice, Inbred NOD
  • Mice, Knockout
  • Mouse Embryonic Stem Cells
  • Mutation
  • Neoplasms, Experimental / drug therapy
  • Neoplasms, Experimental / genetics
  • Neoplasms, Experimental / pathology
  • Phthalazines / pharmacology
  • Phthalazines / therapeutic use
  • Piperazines / pharmacology
  • Piperazines / therapeutic use
  • Topotecan / pharmacology
  • Topotecan / therapeutic use

Substances

  • Antineoplastic Agents
  • BRCA1 Protein
  • LIG4 protein, human
  • Phthalazines
  • Piperazines
  • Topotecan
  • ATM protein, human
  • Ataxia Telangiectasia Mutated Proteins
  • Atm protein, mouse
  • DNA Ligase ATP
  • olaparib