DNA polymerase ε relies on a unique domain for efficient replisome assembly and strand synthesis

Nat Commun. 2020 May 15;11(1):2437. doi: 10.1038/s41467-020-16095-x.

Abstract

DNA polymerase epsilon (Pol ε) is required for genome duplication and tumor suppression. It supports both replisome assembly and leading strand synthesis; however, the underlying mechanisms remain to be elucidated. Here we report that a conserved domain within the Pol ε catalytic core influences both of these replication steps in budding yeast. Modeling cancer-associated mutations in this domain reveals its unexpected effect on incorporating Pol ε into the four-member pre-loading complex during replisome assembly. In addition, genetic and biochemical data suggest that the examined domain supports Pol ε catalytic activity and symmetric movement of replication forks. Contrary to previously characterized Pol ε cancer variants, the examined mutants cause genome hyper-rearrangement rather than hyper-mutation. Our work thus suggests a role of the Pol ε catalytic core in replisome formation, a reliance of Pol ε strand synthesis on a unique domain, and a potential tumor-suppressive effect of Pol ε in curbing genome re-arrangements.

Publication types

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

MeSH terms

  • Cell Cycle
  • Cell Cycle Proteins / genetics
  • Chromosome Structures / genetics
  • DNA Polymerase II / chemistry
  • DNA Polymerase II / physiology*
  • DNA Replication*
  • Doxycycline / pharmacology
  • Gene Expression Regulation*
  • Genome, Human
  • Humans
  • Mutation
  • Neoplasms / genetics
  • Neoplasms / metabolism
  • Poly-ADP-Ribose Binding Proteins / chemistry
  • Poly-ADP-Ribose Binding Proteins / physiology*
  • Protein Domains
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae Proteins / genetics

Substances

  • Cell Cycle Proteins
  • DPB11 protein, S cerevisiae
  • Poly-ADP-Ribose Binding Proteins
  • Saccharomyces cerevisiae Proteins
  • DNA Polymerase II
  • POLE protein, human
  • Doxycycline