Results: 5

1.
FIGURE 4.

FIGURE 4. From: Glutathione Reductase/Glutathione Is Responsible for Cytotoxic Elemental Sulfur Tolerance via Polysulfide Shuttle in Fungi.

Role of GR in oxidative stress responses. A, effects of menadione (MD), diamide (DA), and hydrogen peroxide (H2O2) (1 mm each) on growth of WT and DGR. Strains were incubated on MMEA for 72 h at 30 °C. B, intracellular activities of thioredoxin reductase (TRR), catalase, and cytochrome c peroxidase (CPX). WT and DGR were incubated in MMEA with (+S) or without (−S) 20 mm eq. PS for 24 h at 30 °C.

Ikuo Sato, et al. J Biol Chem. 2011 June 10;286(23):20283-20291.
2.
FIGURE 5.

FIGURE 5. From: Glutathione Reductase/Glutathione Is Responsible for Cytotoxic Elemental Sulfur Tolerance via Polysulfide Shuttle in Fungi.

Sulfur reduction mediated by SR/GR in F. oxysporum and other fungi. A, model of SR/GR-mediated sulfur reduction. B, sulfide production by WT (filled bars) and GR gene disruptants (unfilled bars) of S. cerevisiae and A. nidulans. S. cerevisiae strains BY4741 (WT) and glr1Δ (Δglr1) were used to inoculate medium containing either 20 mm eq. PS or 5 mm eq. CS to optical density of 0.4. The A. nidulans WT and DGR1 (ΔglrA) strains (20) (100 mg dry cells) were cultured in MMEA medium containing 20 mm eq. PS or 5 mm eq. CS for 24 h. C, morphology of S. cerevisiae colonies on MMEA agar plates with or without sodium sulfide, CS, and polysulfide (polyS) (1 mm each) after incubation for 38 h. D, morphology of A. nidulans colonies on MMEA agar plates with or without sodium sulfide, CS, and polysulfide (polyS) (1 mm each) after incubation for 48 h.

Ikuo Sato, et al. J Biol Chem. 2011 June 10;286(23):20283-20291.
3.
FIGURE 1.

FIGURE 1. From: Glutathione Reductase/Glutathione Is Responsible for Cytotoxic Elemental Sulfur Tolerance via Polysulfide Shuttle in Fungi.

Elemental sulfur reduction and toxicity. A, sulfide production by F. oxysporum JCM11502 (WT) incubated in MMEA with various amounts of CS (circles) and PS (triangles) for 24 h at 30 °C. B, numbers of surviving cells after similar incubation with various concentrations of CS (circles), PS (triangles), and sodium sulfide (squares). C, morphology of colonies on MMEA agar plates incubated for 48 h with or without CS and sodium sulfide. D, time-dependent change in sizes of colonies on plates with 5 mm CS (circles), 5 mm sodium sulfide (squares), and no additions (triangles). E and F, effects of thiolate reagents. F. oxysporum WT cells were incubated with or without DTNB, IAA, and IAM (5 mm each) for 2 h, and then sulfide produced within 15 min was monitored (E). Intracellular GSH (filled bars) and GSSG (unfilled bars) were measured in these cells (F). *, less than 1.0 nmol-S mg−1.

Ikuo Sato, et al. J Biol Chem. 2011 June 10;286(23):20283-20291.
4.
FIGURE 2.

FIGURE 2. From: Glutathione Reductase/Glutathione Is Responsible for Cytotoxic Elemental Sulfur Tolerance via Polysulfide Shuttle in Fungi.

Sulfur reductase activities of F. oxysporum. A, cell-free extracts were prepared from F. oxysporum WT (W) and DGR (D) strains cultured with 20 mm eq. PS at 30 °C for 24 h, and rates of sulfide production (left) and NADPH oxidation (right) were determined as described under “Experimental Procedures.” Concentrations of electron mediators and acceptors were set at 1 mm (GSSG), 10 μm (TrxA), 5 mm eq. (CS), and 0.1 mm (polysulfide; polyS). Data are means of three experiments, and error bars represent standard errors. *, less than 0.5 nmol min−1 mg−1. B, reaction catalyzed by SR and polySR. C, amounts of glrA gene transcript determined by quantitative PCR in WT cultured with (+S) or without (−S) 20 mm eq. PS at 30 °C for 12 h. D, enzyme activities of SR (filled bars) and GR (unfilled bars) in WT cultured for 24 h under identical conditions.

Ikuo Sato, et al. J Biol Chem. 2011 June 10;286(23):20283-20291.
5.
FIGURE 3.

FIGURE 3. From: Glutathione Reductase/Glutathione Is Responsible for Cytotoxic Elemental Sulfur Tolerance via Polysulfide Shuttle in Fungi.

Sulfur/glutathione reductase is involved in sulfur tolerance. A, strategy for homologous recombination into glrA locus to construct glrA mutants (left) and Southern blot analysis (right) of WT (lane 1) and DGR (lane 2) strains. Total DNA from strains was digested with EcoRI before blotting and hybridization. B, intracellular GSH and GSSG concentrations. WT and DGR were incubated in MMEA with (+S) or without (−S) 20 mm eq. PS for 24 h at 30 °C. Filled and unfilled bars represent GSH and GSSG, respectively. C, time-dependent production of sulfide by WT (closed) and DGR (open). Strains were cultured in MMEA medium containing 5 mm eq. CS (circles) and 20 mm eq. PS (triangles). Culture flasks were sealed with butyl rubber caps to prevent sulfide evaporation. D, morphology of colonies appeared on MMEA agar plates with or without sodium sulfide, CS, and polysulfide (polyS) (1 mm each) after incubation for 30 h.

Ikuo Sato, et al. J Biol Chem. 2011 June 10;286(23):20283-20291.

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