The production of the reactive oxygen species superoxide and hydrogen peroxide in Saccharomyces cerevisiae induces the expression of various defence genes involved in an oxidative stress response. Expression of many of these genes has been shown to be coordinated by two transcriptional regulators, Yap1p and Skn7p, either alone or in concert. Here, we investigated the role of the Yap1p and Skn7p-mediated stress response in the defence against singlet oxygen, a non-radical reactive oxygen species produced mainly by photosensitized reactions in illuminated cells. Both, a yap1 and skn7 mutant were highly sensitive to Rose Bengal, an exogenous photosensitizer producing singlet oxygen in the light. The expression of a Yap1p-dependent reporter gene was induced by increased singlet oxygen production, showing that singlet oxygen activates general oxidative stress response mechanisms required for the resistance against Rose Bengal treatment. This response was also slightly stimulated by light in the absence of the photosensitizer, possibly due to singlet oxygen production by endogenous photosensitizers. The expression pattern of four oxidative stress genes in a yap1, skn7 and wild-type strain and the sensitivity of the corresponding mutants exposed to different oxidative stress conditions proved a role of Yap1p and Skn7p in the defence against singlet oxygen. Similarities in the genetic responses against singlet oxygen and hydroperoxides suggest an overlap in the oxidative stress response against these reactive oxygen species.
The glutathione peroxidase homologous gene (Gpxh gene) in Chlamydomonas reinhardtii is up-regulated under oxidative stress conditions. The Gpxh gene showed a remarkably strong and fast induction by the singlet oxygen-generating photosensitizers neutral red, methylene blue and rose Bengal. The Gpxh mRNA levels strongly increased, albeit much more slowly, upon exposure to the organic hydroperoxides tert-butyl hydroperoxide (t-BOOH) and cumene hydroperoxide. In contrast, the Gpxh mRNA levels were only weakly induced by exposure to the superoxide-generating compound paraquat and by hydrogen peroxide. A comparison of the Gpxh mRNA levels with those of the heat shock protein HSP70A and the iron superoxide dismutase gene showed qualitative and quantitative differences for the three genes under oxidative stress conditions tested. The Gpxh gene is specifically induced by singlet-oxygen photosensitizers and the relative induction by other compounds is much weaker for Gpxh than for the other genes investigated. Using Gpxh promoter fusions with the arylsulfatase reporter gene, we have shown that the Gpxh was transcriptionally up-regulated by singlet-oxygen photosensitizers. It is also shown that the Gpxh promoter contains a region between 104 and 179 bp upstream of the transcription start that is responsible for the mRNA up-regulation upon exposure to 1O2 but not t-BOOH. Within this region a regulatory sequence homologous to the mammalian cAMP response element (CRE) and activator protein 1 (AP-1) binding site was identified within a 16 bp palindrome.
Exposure of the green alga Chlamydomonas reinhardtii Dangeard to a combination of environmental stress by high light irradiance and chemical stress by each of the three herbicides paraquat, atrazine, and norflurazon resulted in diverse multiple stressor effects on growth and survival of the cells. Under low light conditions, growth analyzed by cell numbers was generally more sensitive to herbicide treatment than optical density-based growth rates or colony-forming unit endpoints, which both also analyzed the viability of the cells. However, growth analyzed by optical density and colony-forming units in herbicide-treated cultures was affected much more strongly by high light irradiance, as shown by reduced 50% effective concentrations, indicating extensive multiple stressor effects of the combined treatment on the viability of the cells. None of the currently used concepts for mixture toxicity (concentration addition, independent action, or effect summation) could accurately describe the effects measured by the two stressors in combination. Both synergistic and antagonistic interactions seem to occur depending on the light conditions and the parameter analyzed. The strong stimulation of toxicity by the combined stresses can be explained by the similar mode of toxic action of the treatments, all increasing the production of reactive oxygen species. Antagonistic effects, conversely, are probably attributable to the various protection mechanisms of photosynthetic organisms to increased light irradiance, which help the cells acclimate to specific light conditions and defend against the deleterious effects of excess light. These protection mechanisms can affect growth and viability under increased light conditions and also might influence the toxicity of the photosynthetic herbicides.
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