Reduced hydroperoxidase (HPI and HPII) activity in the Deltafur mutant contributes to increased sensitivity to UVA radiation in Escherichia coli.

In Escherichia coli, Deltafur (ferric uptake regulator) mutants are hypersensitive to various oxidative agents, including UVA radiation (400-315 nm). Studies suggest that UVA radiation mediates its biological effects on bacteria via oxidative mechanisms that lead to reactive oxygen species, including the superoxide anion radical (O2.-), hydroxyl radical (HO.), hydrogen peroxide (H2O2) and singlet oxygen (1O2). There is accumulating evidence that Fur may play an important role in the defense against UVA radiation. In addition to regulating almost all genes directly involved in iron acquisition, Fur also regulates the expression of manganese and iron superoxide dismutase (MnSOD, FeSOD), key enzymes in the defense against oxygen toxicity in E. coli. In Deltafur mutants, there is a complete absence of FeSOD. Previous results suggest that the native iron chelating agent, enterobactin, which exists in increased levels in Deltafur mutants, is an endogenous chromophore for UVA, releasing Fe2+ into the cytoplasm to catalyze the production of highly reactive hydroxyl radicals. We now report that the hypersensitivity of Deltafur mutants to UVA irradiation is associated with reduced hydroperoxidase I (HPI) and hydroperoxidase II (HPII) activity, and is associated with a decrease in the transcription of katE and katG genes. The observed decrease in HPII activity in Deltafur mutants is also associated with reduced rpoS gene transcription. This study provides additional evidence that the Fur gene product, in addition to its known regulatory effect on the expression of SOD and iron uptake mechanisms, also regulates HPI and HPII activity levels in E. coli. An H2O2-inducible antioxidant defense system leading to an increase in HPI activity, is unaltered in Deltafur mutants.

[1]  Roberto Kolter,et al.  The dps promoter is activated by OxyR during growth and by IHF and σs in stationary phase , 1994, Molecular microbiology.

[2]  P. Loewen,et al.  Regulation of transcription of katE and katF in Escherichia coli , 1990, Journal of bacteriology.

[3]  G. Storz,et al.  Transcriptional regulator of oxidative stress-inducible genes: direct activation by oxidation. , 1990, Science.

[4]  P. Loewen,et al.  Genetic mapping of katF, a locus that with katE affects the synthesis of a second catalase species in Escherichia coli , 1984, Journal of bacteriology.

[5]  D. Häder,et al.  Involvement of reactive oxygen species in the UV-B damage to the cyanobacterium Anabaena sp. , 2002, Journal of photochemistry and photobiology. B, Biology.

[6]  Jeffrey H. Miller Experiments in molecular genetics , 1972 .

[7]  D. Touati,et al.  Mutations by near-ultraviolet radiation in Escherichia coli strains lacking superoxide dismutase. , 1989, Mutation research.

[8]  G. F. Kramer,et al.  Oxidative mechanisms of toxicity of low-intensity near-UV light in Salmonella typhimurium , 1987, Journal of bacteriology.

[9]  K. Hantke,et al.  Fur regulon in gram-negative bacteria. Identification and characterization of new iron-regulated Escherichia coli genes by a fur titration assay. , 1994, Journal of molecular biology.

[10]  M. Schuler,et al.  Gene regulation by low level UV-B radiation: identification by DNA array analysis , 2002, Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology.

[11]  Sarah Dubrac,et al.  Fur Positive Regulation of Iron Superoxide Dismutase in Escherichia coli: Functional Analysis of thesodB Promoter , 2000, Journal of bacteriology.

[12]  D. Hassett,et al.  Ferric uptake regulator (Fur) mutants of Pseudomonas aeruginosa demonstrate defective siderophore-mediated iron uptake, altered aerobic growth, and decreased superoxide dismutase and catalase activities , 1996, Journal of bacteriology.

[13]  L. Schrum,et al.  The effects of fur on the transcriptional and post-transcriptional regulation of MnSOD gene (sodA) in Escherichia coli. , 1994, Archives of biochemistry and biophysics.

[14]  Thomas D. Schneider,et al.  OxyR and SoxRS Regulation offur , 1999, Journal of bacteriology.

[15]  Chris E Cooper,et al.  Global Iron-dependent Gene Regulation in Escherichia coli , 2003, Journal of Biological Chemistry.

[16]  J. Fee,et al.  Control of Escherichia coli superoxide dismutase (sodA and sodB) genes by the ferric uptake regulation (fur) locus , 1990, Journal of bacteriology.

[17]  G. Glinsky,et al.  Role of rpoS (katF) in oxyR‐independent regulation of hydroperoxidase I in Escherichia coli , 1994, Molecular microbiology.

[18]  I. Fridovich,et al.  Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. , 1971, Analytical biochemistry.

[19]  H. Schellhorn,et al.  Transcriptional regulation of katE in Escherichia coli K-12 , 1988, Journal of bacteriology.

[20]  P. Loewen,et al.  Catalases HPI and HPII in Escherichia coli are induced independently. , 1985, Archives of biochemistry and biophysics.

[21]  M. A. Stahmann,et al.  An improved procedure using ferricyanide for detecting catalase isozymes. , 1971, Analytical biochemistry.

[22]  S. Clarke,et al.  RpoS- and OxyR-independent induction of HPI catalase at stationary phase in Escherichia coli and identification of rpoS mutations in common laboratory strains , 1997, Journal of bacteriology.

[23]  A. Eisenstark,et al.  Role of Enterobactin and Intracellular Iron in Cell Lethality During Near‐UV Irradiation in Escherichia coli , 1996, Photochemistry and photobiology.

[24]  R. Hengge-aronis,et al.  Signal Transduction and Regulatory Mechanisms Involved in Control of the σS (RpoS) Subunit of RNA Polymerase , 2002, Microbiology and Molecular Biology Reviews.

[25]  I. Fridovich,et al.  Effects of molecular oxygen on detection of superoxide radical with nitroblue tetrazolium and on activity stains for catalase. , 1984, Analytical biochemistry.

[26]  V. Braun Avoidance of iron toxicity through regulation of bacterial iron transport. , 1997, Biological chemistry.

[27]  D. Touati,et al.  Lethal oxidative damage and mutagenesis are generated by iron in delta fur mutants of Escherichia coli: protective role of superoxide dismutase , 1995, Journal of bacteriology.

[28]  D. Touati,et al.  Two global regulators repress the anaerobic expression of MnSOD in Escherichia coli: Fur (ferric uptake regulation) and Arc (aerobic respiration control) , 1991, Molecular microbiology.