Okazaki fragment maturation involves α-segment error editing by the mammalian FEN1/MutSα functional complex

EMBO J. 2015 Jul 2;34(13):1829-43. doi: 10.15252/embj.201489865. Epub 2015 Apr 28.

Abstract

During nuclear DNA replication, proofreading-deficient DNA polymerase α (Pol α) initiates Okazaki fragment synthesis with lower fidelity than bulk replication by proofreading-proficient Pol δ or Pol ε. Here, we provide evidence that the exonuclease activity of mammalian flap endonuclease (FEN1) excises Pol α replication errors in a MutSα-dependent, MutLα-independent mismatch repair process we call Pol α-segment error editing (AEE). We show that MSH2 interacts with FEN1 and facilitates its nuclease activity to remove mismatches near the 5' ends of DNA substrates. Mouse cells and mice encoding FEN1 mutations display AEE deficiency, a strong mutator phenotype, enhanced cellular transformation, and increased cancer susceptibility. The results identify a novel role for FEN1 in a specialized mismatch repair pathway and a new cancer etiological mechanism.

Keywords: DNA mismatch repair; MutSα; Okazaki fragment maturation; flap endonuclease 1; α‐segment error editing.

Publication types

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

MeSH terms

  • Animals
  • Cells, Cultured
  • DNA / metabolism*
  • DNA Mismatch Repair* / genetics
  • DNA Polymerase I / metabolism*
  • DNA Replication / genetics
  • Embryo, Mammalian
  • Female
  • Flap Endonucleases / classification
  • Flap Endonucleases / genetics
  • Flap Endonucleases / metabolism*
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Male
  • Mice
  • Mice, Transgenic
  • MutS DNA Mismatch-Binding Protein / genetics
  • MutS DNA Mismatch-Binding Protein / metabolism*
  • Saccharomyces cerevisiae

Substances

  • Okazaki fragments
  • DNA
  • DNA Polymerase I
  • Flap Endonucleases
  • MutS DNA Mismatch-Binding Protein