Resistance to nitrophenolic herbicides and metronidazole in the cyanobacterium Synechocystis sp. PCC 6803 as a result of the inactivation of a nitroreductase-like protein encoded by drgA gene.
Dinoseb is a herbicide known to inhibit photosystem II electron transfer like DCMU, triazine and phenolic-type herbicides. The mutant Din7 of the cyanobacterium Synechocystis sp. PCC 6803, selected for resistance to dinoseb, and the mutant Ins2, constructed by the insertion of the kanamycin resistance cassette into the drgA gene, were cross-resistant to other nitrophenolic herbicides (DNOC, 2,4-dinitrophenol) and to the cell inhibitor metronidazole but not to the photosystem II inhibitors DCMU or ioxynil. The Din7 mutant had the same characteristics of photosystem II inhibition by dinoseb as the wild type. This result suggested the existence of another site for dinoseb inhibition. The wild type cells modified dinoseb to a non-toxic product that gave an absorption spectrum similar to that of dithionite treated dinoseb containing reduced nitro groups. In contrast, the Din7 mutant did not modify dinoseb. These phenomena were controlled by the drgA gene encoding a protein which showed similarity to several enzymes having nitroreductase activity. The addition of superoxide dismutase to the medium relieved the toxic effect of dinoseb in wild type cells but not in Din7. It is proposed that in wild type cells of Synechocystis sp. PCC 6803 the DrgA protein is involved in detoxification of dinoseb via the reduction of the nitro group(s) and this process is accompanied by the formation of toxic superoxide anions. Mutations blocking the activity of the DrgA protein lead to the development of resistance to nitrophenolic herbicides and metronidazole.
Elanskaya
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Department of Genetics, Moscow State University, Russia. irina@selansky.home.bio.msu.ru
Chesnavichene
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EA
Vernotte
C
C
Astier
C
C
D019297:2,4-Dinitrophenol / Q000031:analogs & derivatives / Q000494:pharmacology; D000595:Amino Acid Sequence; D000458:Cyanobacteria / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics; D004138:Dinitrocresols / Q000494:pharmacology; D004352:Drug Resistance, Microbial; D005798:Genes, Bacterial; D006540:Herbicides / Q000494:pharmacology*; D007700:Kinetics; D008795:Metronidazole / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009596:Nitrophenols / Q000494:pharmacology*; D009601:Nitroreductases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D045322:Photosynthetic Reaction Center Complex Proteins / Q000187:drug effects / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013053:Spectrophotometry
D004138:Dinitrocresols; D006540:Herbicides; D009596:Nitrophenols; D045322:Photosynthetic Reaction Center Complex Proteins; D008795:Metronidazole; C024132:4,6-dinitro-o-cresol; D009601:Nitroreductases; D019297:2,4-Dinitrophenol; C018527:dinoseb