DNA polymerase IV mediates efficient and quick recovery of replication forks stalled at N2-dG adducts

Nucleic Acids Res. 2014 Jul;42(13):8461-72. doi: 10.1093/nar/gku547. Epub 2014 Jun 23.

Abstract

Escherichia coli DNA polymerase IV (Pol IV, also known as DinB) is a Y-family DNA polymerase capable of catalyzing translesion DNA synthesis (TLS) on certain DNA lesions, and accumulating data suggest that Pol IV may play an important role in copying various kinds of spontaneous DNA damage including N(2)-dG adducts and alkylated bases. Pol IV has a unique ability to coexist with Pol III on the same β clamp and to positively dissociate Pol III from β clamp in a concentration-dependent manner. Reconstituting the entire process of TLS in vitro using E. coli replication machinery and Pol IV, we observed that a replication fork stalled at (-)-trans-anti-benzo[a]pyrene-N(2)-dG lesion on the leading strand was efficiently and quickly recovered via two sequential switches from Pol III to Pol IV and back to Pol III. Our results suggest that TLS by Pol IV smoothes the way for the replication fork with minimal interruption.

Publication types

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

MeSH terms

  • Benzopyrenes*
  • DNA / biosynthesis
  • DNA Adducts*
  • DNA Polymerase III / metabolism
  • DNA Polymerase beta / metabolism*
  • DNA Replication*
  • Deoxyguanosine / analogs & derivatives*
  • Escherichia coli / genetics
  • Escherichia coli Proteins / metabolism*

Substances

  • Benzopyrenes
  • DNA Adducts
  • DinB protein, E coli
  • Escherichia coli Proteins
  • benzo(a)pyrene N2-dG adduct
  • DNA
  • DNA Polymerase III
  • DNA Polymerase beta
  • Deoxyguanosine