Transcriptional regulation by iron of genes encoding iron- and manganese-superoxide dismutases from Pseudomonas putida.
Genes from Pseudomonas putida (Pp), sodA, encoding manganese-superoxide dismutase (MnSOD) and, sodB, iron-superoxide dismutase (FeSOD) were cloned by hybridization with digoxigenin (dig)-labeled PCR products generated from Pp genomic DNA. The sodB gene had a 594 bp open reading frame (ORF), corresponding to 198 amino acids (aa), and a transcript of 880 bases. The sodA gene contained a 609 bp ORF encoding 203 aa and was transcribed as part of a polycistronic operon, consisting of orfY-fumC-orfX-sodA. Pp sodA or sodB genes both restored aerobic growth, growth on paraquat, and growth on minimal medium to an Escherichia coli (Ec) mutant deficient in SOD activity. Paraquat treatment did not enhance mRNA transcription of the sod genes or increase SOD activity in Pp. The Pp sodB gene was highly expressed throughout logarithmic-(log) growth phase and stationary-phase cells grown in medium supplemented with FeCl3, but was down-regulated in iron-deficient conditions, such as in stationary-phase or generated by 2,2'-dipyridyl (DP) treatment. This is the first evidence that iron regulates expression of the sodB gene at the transcriptional level. In contrast, iron-deficient conditions, or addition of MnCl2 to the growth medium, induced transcripts (2.4 kb and 1.2 kb) from the sodA operon. Our results reveal an intricate role of iron in the transcriptional regulation of both Pp sodA and sodB genes.
Kim
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YC
Department of Biology, Utah State University, Logan 84322-5305, USA.
Miller
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CD
Anderson
A J
AJ
D002455:Cell Division / Q000187:drug effects; D003001:Cloning, Molecular; D003470:Culture Media / Q000494:pharmacology; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D005816:Genetic Complementation Test; D007501:Iron / Q000494:pharmacology*; D007527:Isoenzymes / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D010269:Paraquat / Q000494:pharmacology; D016958:Pseudomonas putida / Q000166:cytology / Q000201:enzymology / Q000235:genetics*; D017422:Sequence Analysis, DNA; D013482:Superoxide Dismutase / Q000235:genetics*; D014158:Transcription, Genetic / Q000187:drug effects
D003470:Culture Media; D004269:DNA, Bacterial; D007527:Isoenzymes; D007501:Iron; D013482:Superoxide Dismutase; D010269:Paraquat