Genetic determinants of cellular addiction to DNA polymerase theta

Nat Commun. 2019 Sep 19;10(1):4286. doi: 10.1038/s41467-019-12234-1.

Abstract

Polymerase theta (Pol θ, gene name Polq) is a widely conserved DNA polymerase that mediates a microhomology-mediated, error-prone, double strand break (DSB) repair pathway, referred to as Theta Mediated End Joining (TMEJ). Cells with homologous recombination deficiency are reliant on TMEJ for DSB repair. It is unknown whether deficiencies in other components of the DNA damage response (DDR) also result in Pol θ addiction. Here we use a CRISPR genetic screen to uncover 140 Polq synthetic lethal (PolqSL) genes, the majority of which were previously unknown. Functional analyses indicate that Pol θ/TMEJ addiction is associated with increased levels of replication-associated DSBs, regardless of the initial source of damage. We further demonstrate that approximately 30% of TCGA breast cancers have genetic alterations in PolqSL genes and exhibit genomic scars of Pol θ/TMEJ hyperactivity, thereby substantially expanding the subset of human cancers for which Pol θ inhibition represents a promising therapeutic strategy.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Aminoquinolines / toxicity
  • Animals
  • Breast Neoplasms / genetics*
  • CRISPR-Cas Systems / genetics
  • Cell Line
  • DNA Breaks, Double-Stranded
  • DNA End-Joining Repair / genetics*
  • DNA Polymerase theta
  • DNA-Directed DNA Polymerase / genetics*
  • DNA-Directed DNA Polymerase / metabolism
  • HEK293 Cells
  • Humans
  • Mice
  • Mitomycin / toxicity
  • Picolinic Acids / toxicity

Substances

  • Aminoquinolines
  • Picolinic Acids
  • pyridostatin
  • Mitomycin
  • DNA-Directed DNA Polymerase