Mutagenicity of p-aminophenol in E. coli WP2uvrA/pKM101 and its relevance to oxidative DNA damage.
It was recently reported that p-aminophenol (p-AP) induces DNA cleavage in mouse lymphoma cells, CHO cells and human lymphoblastoid cells. The mutagenicity of p-AP has not, however, been detected by reverse mutation assays. The purpose of this study was to assess the mutagenicity of p-AP by reverse mutation assay using Escherichia coli WP2uvrA/pKM101, which has a spectrum for detecting mutations different from those of other strains in the family with an AT base pair at the mutation site and has higher sensitivity to certain oxidative mutagens as compared to other strains. We found that p-AP was mutagenic to E. coli WP2uvrA/pKM101. The mutagenic activity of this compound was suppressed with the addition of dimethylsulfoxide or catalase, suggesting the involvement of active oxygen species in the mutagenic process induced by p-AP. To further elucidate the underlying mechanism, we used isolated DNA for the following experiments. It was revealed, by gel electrophoretic analysis, that p-AP induced DNA cleavage in the presence of Fe(III). However, p-AP alone did not induce this cleavage. Formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine by p-AP in calf thymus DNA was also detected in the presence of Fe(III) by HPLC with an electrochemical detector. ESR-spin trapping experiments using DMPO detected the production of hydroxyl radical (.OH) in the solution of p-AP with Fe(III). Both p-AP mediated DNA damages and .OH production by p-AP in the presence of Fe(III) were completely inhibited by .OH scavengers (ethanol, mannitol, sodium formate, dimethylsulfoxide) and catalase. These results suggest that .OH derived from the reaction between H2O2 and Fe(III) (Fenton reaction) participates in the oxidative DNA damage. Accordingly, the same mechanism might be working in E. coli WP2uvrA/pKM101 during induction of the mutation by p-AP.
Department of Public Health and Environmental Medicine, Jikei University School of Medicine, Tokyo, Japan.
R
R
Yoshida
S
S
Oikawa
Y
Y
Ogawa
Y
Y
Miyakoshi
M
M
Ooida
K
K
Asanuma
H
H
Shimizu
D000251:Adenosine Triphosphatases / Q000235:genetics; D000627:Aminophenols / Q000633:toxicity*; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D002374:Catalase / Q000378:metabolism; D002417:Cattle; D004249:DNA Damage; D004269:DNA, Bacterial / Q000187:drug effects; D004268:DNA-Binding Proteins / Q000235:genetics; D004926:Escherichia coli / Q000187:drug effects; D029968:Escherichia coli Proteins; D005290:Ferric Compounds; D016166:Free Radical Scavengers; D017665:Hydroxyl Radical; D009152:Mutagenicity Tests; D009153:Mutagens / Q000633:toxicity*; D010084:Oxidation-Reduction; D014466:Ultraviolet Rays
D000627:Aminophenols; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D005290:Ferric Compounds; D016166:Free Radical Scavengers; D009153:Mutagens; D017665:Hydroxyl Radical; D002374:Catalase; C068631:UvrA protein, E coli; D000251:Adenosine Triphosphatases; C026729:4-aminophenol