Involvement of DnaE, the second replicative DNA polymerase from Bacillus subtilis, in DNA mutagenesis

J Biol Chem. 2004 Jan 16;279(3):1757-67. doi: 10.1074/jbc.M310719200. Epub 2003 Oct 30.

Abstract

In a large group of organisms including low G + C bacteria and eukaryotic cells, DNA synthesis at the replication fork strictly requires two distinct replicative DNA polymerases. These are designated pol C and DnaE in Bacillus subtilis. We recently proposed that DnaE might be preferentially involved in lagging strand synthesis, whereas pol C would mainly carry out leading strand synthesis. The biochemical analysis of DnaE reported here is consistent with its postulated function, as it is a highly potent enzyme, replicating as fast as 240 nucleotides/s, and stalling for more than 30 s when encountering annealed 5'-DNA end. DnaE is devoid of 3' --> 5'-proofreading exonuclease activity and has a low processivity (1-75 nucleotides), suggesting that it requires additional factors to fulfill its role in replication. Interestingly, we found that (i) DnaE is SOS-inducible; (ii) variation in DnaE or pol C concentration has no effect on spontaneous mutagenesis; (iii) depletion of pol C or DnaE prevents UV-induced mutagenesis; and (iv) purified DnaE has a rather relaxed active site as it can bypass lesions that generally block other replicative polymerases. These results suggest that DnaE and possibly pol C have a function in DNA repair/mutagenesis, in addition to their role in DNA replication.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus subtilis / enzymology*
  • Bacterial Proteins*
  • DNA Adducts / metabolism
  • DNA Polymerase III / physiology*
  • DNA Repair
  • DNA Replication*
  • DNA-Directed DNA Polymerase / physiology
  • Mutagenesis*
  • SOS Response, Genetics

Substances

  • Bacterial Proteins
  • DNA Adducts
  • DNA polymerase III, alpha subunit
  • PolC protein, bacteria
  • DNA Polymerase III
  • DNA-Directed DNA Polymerase