Roles of E. Coli DNA Polymerases IV and V in Lesion-Targeted and Untargeted SOS Mutagenesis

Nature. 2000 Apr 27;404(6781):1014-8. doi: 10.1038/35010020.

Abstract

The expression of the Escherichia coli DNA polymerases pol V (UmuD'2C complex) and pol IV (DinB) increases in response to DNA damage. The induction of pol V is accompanied by a substantial increase in mutations targeted at DNA template lesions in a process called SOS-induced error-prone repair. Here we show that the common DNA template lesions, TT (6-4) photoproducts, TT cis-syn photodimers and abasic sites, are efficiently bypassed within 30 seconds by pol V in the presence of activated RecA protein (RecA*), single-stranded binding protein (SSB) and pol III's processivity beta,gamma-complex. There is no detectable bypass by either pol IV or pol III on this time scale. A mutagenic 'signature' for pol V is its incorporation of guanine opposite the 3'-thymine of a TT (6-4) photoproduct, in agreement with mutational spectra. In contrast, pol III and pol IV incorporate adenine almost exclusively. When copying undamaged DNA, pol V exhibits low fidelity with error rates of around 10(-3) to 10(-4), with pol IV being 5- to 10-fold more accurate. The effects of RecA protein on pol V, and beta,gamma-complex on pol IV, cause a 15,000- and 3,000-fold increase in DNA synthesis efficiency, respectively. However, both polymerases exhibit low processivity, adding 6 to 8 nucleotides before dissociating. Lesion bypass by pol V does not require beta,gamma-complex in the presence of non-hydrolysable ATPgammaS, indicating that an intact RecA filament may be required for translesion synthesis.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • DNA Damage*
  • DNA Polymerase beta / metabolism*
  • DNA, Bacterial / biosynthesis
  • DNA, Bacterial / metabolism
  • DNA-Directed DNA Polymerase / metabolism*
  • Escherichia coli / enzymology*
  • Escherichia coli / genetics
  • Escherichia coli Proteins
  • Mutagenesis
  • Rec A Recombinases / metabolism
  • SOS Response, Genetics*
  • Templates, Genetic

Substances

  • DNA, Bacterial
  • Escherichia coli Proteins
  • DNA Polymerase beta
  • Rec A Recombinases
  • DNA polymerase V, E coli
  • DNA-Directed DNA Polymerase