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Modulation of the H2O2-induced SOS response in Escherichia coli PQ300 by amino acids, metal chelators, antioxidants, and scavengers of reactive oxygen species. Environ Mol Mutagen 1993;22(3):157-63

Date

01/01/1993

Pubmed ID

8404875

DOI

10.1002/em.2850220308

Scopus ID

2-s2.0-0027331518   22 Citations

Abstract

The SOS chromotest is a simple colorimetric genotoxicity assay that monitors DNA repair by measuring the induction of the gene sfiA in Escherichia coli K-12. E. coli PQ300, a diagnostic SOS tester strain for the detection of oxidative genotoxins, carries a mutation in a key gene for antioxidative defense, oxyR. This mutation renders PQ300 more sensitive to oxidative genotoxins, particularly to H2O2. We found that induction of the SOS response by H2O2 in E. coli PQ300 is dependent on the composition of the incubation medium; a substantially reduced response was obtained in minimal phosphate buffered saline (PBS) as opposed to complex Luria broth (LB) medium. Supplementation of PBS with histidine or cysteine stimulated H2O2-induced SOS induction to levels exceeding those found in LB medium. Low concentrations of glutathione (20-70 microM) also enhanced the H2O2-induced SOS response in E. coli PQ300, whereas higher concentrations (> 150 microM) were protective. Preincubation of tester cells with the chelators o-phenanthroline, 2,2-dipyridyl, and ethylenediaminetetraacetic acid (EDTA) protected cells from the effects of H2O2, although EDTA was only partially effective. Pretreatment of PQ300 with the antioxidant ascorbic acid or the hydroxyl radical scavenger dimethyl sulfoxide also diminished the SOS response, whereas mannitol and glucose were ineffective. The results show that the net effect of H2O2-induced DNA damage is influenced by the balance of oxidative and antioxidative factors and, furthermore, can be modulated by constituents of the extracellular milieu.

Author List

Müller J, Janz S

Author

Siegfried Janz MD Professor in the Medicine department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Amino Acids
Antimutagenic Agents
Antioxidants
Chelating Agents
Culture Media
DNA Damage
Escherichia coli
Free Radical Scavengers
Hydrogen Peroxide
Mutagenicity Tests
Mutagens
Oxidation-Reduction
Phagocytes
Reactive Oxygen Species