Role of DnaB helicase in UV-induced illegitimate recombination in Escherichia coli

J Bacteriol. 2001 Sep;183(17):4964-9. doi: 10.1128/JB.183.17.4964-4969.2001.

Abstract

To study the involvement of DNA replication in UV-induced illegitimate recombination, we examined the effect of temperature-sensitive dnaB mutations on illegitimate recombination and found that the frequency of illegitimate recombination was reduced by an elongation-deficient mutation, dnaB14, but not by an initiation-deficient mutation, dnaB252. This result indicates that DNA replication is required for UV-induced illegitimate recombination. In addition, the dnaB14 mutation also affected spontaneous or UV-induced illegitimate recombination enhanced by the recQ mutation. Nucleotide sequence analyses of the recombination junctions showed that DnaB-mediated illegitimate recombination is short homology dependent. Previously, Michel et al. (B. Michel, S. Ehrlich, and M. Uzest, EMBO J. 16:430--438, 1997) showed that thermal treatment of the temperature-sensitive dnaB8 mutant induces double-stranded breaks, implying that induction of illegitimate recombination occurs. To explain the discrepancy between the observations, we propose a model for DnaB function, in which the dnaB mutations may exhibit two types of responses, early and late responses, for double-stranded break formation. In the early response, replication forks stall at damaged DNA, resulting in the formation of double-stranded breaks, and the dnaB14 mutation reduces the double-stranded breaks shortly after temperature shift-up. On the other hand, in the late response, the arrested replication forks mediated by the dnaB8 mutation may induce double-stranded breaks after prolonged incubation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins*
  • DNA Damage
  • DNA Helicases / genetics
  • DNA Helicases / physiology*
  • DNA, Bacterial / chemistry
  • DnaB Helicases
  • Escherichia coli / enzymology
  • Escherichia coli / genetics*
  • Escherichia coli / radiation effects
  • Mutation
  • Recombination, Genetic / radiation effects*
  • Sequence Analysis, DNA
  • Ultraviolet Rays*

Substances

  • Bacterial Proteins
  • DNA, Bacterial
  • DNA Helicases
  • DnaB Helicases