Mismatch repair operates at the replication fork in direct competition with mismatch extension by DNA polymerase δ

Roland Klassen, Venkat Gangavarapu, Robert E. Johnson, Louise Prakash, Satya Prakash

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

DNA mismatch repair (MMR) in eukaryotes is believed to occur post-replicatively, wherein nicks or gaps in the nascent DNA strand are suggested to serve as strand discrimination signals. However, how such signals are generated in the nascent leading strand has remained unclear. Here we examine the alternative possibility that MMR occurs in conjunction with the replication fork. To this end, we utilize mutations in the PCNA interacting peptide (PIP) domain of the Pol3 or Pol32 subunit of DNA polymerase δ (Polδ) and show that these pip mutations suppress the greatly elevated mutagenesis in yeast strains harboring the pol3-01 mutation defective in Polδ proofreading activity. And strikingly, they suppress the synthetic lethality of pol3-01 pol2-4 double mutant strains, which arises from the vastly enhanced mutability due to defects in the proofreading functions of both Polδ and Polε. Our finding that suppression of elevated mutagenesis in pol3-01 by the Polδ pip mutations requires intact MMR supports the conclusion that MMR operates at the replication fork in direct competition with other mismatch removal processes and with extension of synthesis from the mispair by Polδ. Furthermore, the evidence that Polδ pip mutations eliminate almost all the mutability of pol2-4 msh2Δ or pol3-01 pol2-4 adds strong support for a major role of Polδ in replication of both the leading and lagging DNA strands.

Original languageEnglish (US)
Article number104598
JournalJournal of Biological Chemistry
Volume299
Issue number4
DOIs
StatePublished - Apr 2023

Keywords

  • DNA polymerase delta
  • DNA polymerase epsilon
  • MMR at replication fork
  • PCNA
  • PCNA interacting domains in Pol delta
  • mismatch repair

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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