Stationary-phase mutation in the bacterial chromosome: recombination protein and DNA polymerase IV dependence

Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8334-41. doi: 10.1073/pnas.151009798.

Abstract

Several microbial systems have been shown to yield advantageous mutations in slowly growing or nongrowing cultures. In one assay system, the stationary-phase mutation mechanism differs from growth-dependent mutation, demonstrating that the two are different processes. This system assays reversion of a lac frameshift allele on an F' plasmid in Escherichia coli. The stationary-phase mutation mechanism at lac requires recombination proteins of the RecBCD double-strand-break repair system and the inducible error-prone DNA polymerase IV, and the mutations are mostly -1 deletions in small mononucleotide repeats. This mutation mechanism is proposed to occur by DNA polymerase errors made during replication primed by recombinational double-strand-break repair. It has been suggested that this mechanism is confined to the F plasmid. However, the cells that acquire the adaptive mutations show hypermutation of unrelated chromosomal genes, suggesting that chromosomal sites also might experience recombination protein-dependent stationary-phase mutation. Here we test directly whether the stationary-phase mutations in the bacterial chromosome also occur via a recombination protein- and pol IV-dependent mechanism. We describe an assay for chromosomal mutation in cells carrying the F' lac. We show that the chromosomal mutation is recombination protein- and pol IV-dependent and also is associated with general hypermutation. The data indicate that, at least in these male cells, recombination protein-dependent stationary-phase mutation is a mechanism of general inducible genetic change capable of affecting genes in the bacterial chromosome.

Publication types

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

MeSH terms

  • Alleles
  • Bacterial Proteins / genetics
  • Chromosomes, Bacterial*
  • DNA Polymerase beta / metabolism*
  • Endodeoxyribonucleases / genetics
  • Escherichia coli Proteins*
  • Exodeoxyribonuclease V
  • Exodeoxyribonucleases / genetics
  • Genes, Bacterial
  • Lactose / metabolism
  • Mutagenesis
  • Rec A Recombinases / genetics
  • Recombination, Genetic*
  • Repressor Proteins / metabolism

Substances

  • Bacterial Proteins
  • Escherichia coli Proteins
  • Repressor Proteins
  • ruvC protein, E coli
  • tetracycline resistance-encoding transposon repressor protein
  • RecG protein, E coli
  • Rec A Recombinases
  • DNA Polymerase beta
  • Endodeoxyribonucleases
  • Exodeoxyribonucleases
  • Exodeoxyribonuclease V
  • Lactose