PMID:16824196

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Citation

Sakai, A, Nakanishi, M, Yoshiyama, K and Maki, H (2006) Impact of reactive oxygen species on spontaneous mutagenesis in Escherichia coli.Genes Cells 11:767-78

Abstract

Reactive oxygen species (ROS) are potent oxidants that attack chromosomal DNA and free nucleotides, leading to oxidative DNA damage that causes genetic alterations. To avoid the ROS-mediated mutagenesis, cells have elaborate mechanisms including powerful antioxidant components and repair pathways that eliminate oxidative DNA damage. Because of the effective anti-mutagenic functions, it has been unclear to what extent the ROS contribute to spontaneous mutagenesis. Here we show that a significant portion of spontaneous mutations is actually caused by the ROS in aerobically growing Escherichia coli cells. Using the rpsL gene as a mutational target sequence, we established an experimental procedure to analyze spontaneous mutations occurring under a strictly anaerobic condition. Strong mutator phenotypes of cells defective in both mutM and mutY genes or ones lacking mutT gene were completely suppressed under the anaerobic condition, indicative of an absence of hydroxyl radicals in the cells. From a series of analyses with wild-type E. coli cells grown under different redox conditions, it appeared that 89% of base substitutions were caused by the ROS, especially hydroxyl radicals, in cells growing in the atmosphere. The ROS-mediated spontaneous mutations included highly site-specific base substitutions, two types of randomly occurring transversions, G:C-->C:G and A:T-->T:A, and -1 frameshifts at non-iterated base sequences.

Links

PubMed Online version:10.1111/j.1365-2443.2006.00982.x

Keywords

Aerobiosis; Anaerobiosis; Base Sequence; Escherichia coli; Escherichia coli Proteins; Hydrogen Peroxide; Hydroxyl Radical; Molecular Sequence Data; Mutation; Reactive Oxygen Species; Ribosomal Proteins

Main Points of the Paper

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Materials and Methods Used

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Phenotype Annotations

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Phenotype of Taxon Information Genotype Information (if known) Condition Information OMP ID OMP Term Name ECO ID ECO Term Name Notes Status

a mutation or genetic difference within a strain

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: MK6168
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain: fur+
  • Genotype of Experimental Strain : fur(del)::kan
  • Reference Condition:
OMP:0005068

increased mutation frequency

Figure 4: aerobic growth conditions.

MK6168 = MK811 fur(del)::kan The fur(del)::kan allele was transduced from QC1732.[1]

a single strain under different conditions

  • Taxon: Escherichia coli
  • Strain: K-12
  • Substrain: MK811
  • NCBI Taxon ID: 83333
  • Genotype of Reference Strain:
  • Reference Condition: aerobic
  • Experimental Condition: anaerobic
OMP:0005069

decreased mutation frequency

Figure 1: decreased frequency of base substitution mutations

MK811 = MG1655 cysE::rpsL (735 bp DNA fragment containing rpsL+)



Notes

References

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  1. Nunoshiba, T et al. (1999) Role of iron and superoxide for generation of hydroxyl radical, oxidative DNA lesions, and mutagenesis in Escherichia coli. J. Biol. Chem. 274 34832-7 PubMed OMPwiki page