A novel test for identifying genes involved in aldehyde detoxification in the yeast. Increased sensitivity of superoxide‐deficient yeast to aldehydes and their metabolic precursors

Abstract

A novel test for the identification of genes involved in aldehyde metabolism is proposed, based on detection of altered sensitivity of the yeast to corresponding alcohols, metabolic precursors of the aldehydes. This attitude enabled to an unexpected detection increased sensitivity of mutants devoid of CuZn-superoxide dismutase (CuZnSOD) to allyl alcohol (precursor of acrolein) and nonenol. We interpret this finding as due to inactivation of some important element of aldehyde detoxification by increased flux of superoxide in DeltaCuZnSOD mutants.

Acrolein toxicity involves oxidative stress caused by glutathione depletion in the yeast Saccharomyces cerevisiae

“…The present results show that wild-type strains of various genetic background differ in the sensitivity to allyl alcohol, confirming our previous data on the hypersensitivity of Δsod1 mutant to allyl alcohol (Biliński et al 2005) and demonstrate that mutants defective in other elements of the antioxidant barrier (devoid of the Yap1p transcription factor, of thioredoxins 1+2, and deficient in the synthesis of glutathione) are also hypersensitive to allyl alcohol (example of the sensitivity drop test is shown in Fig. 1, summary in Table 2).…”

“…Colony growth was inspected after 48 h determined the intracellular level of glutathione. Depletion of acid-soluble cellular thiols by exposure of yeast cells to allyl alcohol has been reported by us previously (Biliński et al 2005). In this study, we confirm this finding using cell staining with monochlorobimane, which shows a high degree of specificity for glutathione (Nair et al 1991) demonstrating a decrease in cell staining with this probe with increasing concentration of allyl alcohol (Fig.…”

“…The way of growth assessment was the same as in Table 2. ADH is able to oxidize allyl alcohol and ethanol (Leskovac et al 2002), and exposure of the yeast to allyl alcohol leads to a considerable increase in the aldehyde content of the cells (Biliński et al 2005). Pyrazole, a competitive inhibitor of ADH (Leskovac et al 1998), prevented the toxicity of allyl alcohol (Fig.…”

“…Acrolein toxicity has been suggested not to be mediated by generation of reactive oxygen species (ROS), as no genes coding for antioxidant molecules have been found to be important for the resistance to acrolein (Trotter et al 2006). On the other hand, hypersensitivity of yeast mutants lacking the Yap1 transcriptional activator protein or defective in glutathione (GSH) metabolism to malondialdehyde and lipid hydroperoxides has been demonstrated (Turton et al 1997), and our previous studies have revealed increased sensitivity of superoxide dismutase-deficient yeast to acrolein (Biliński et al 2005) and other aldehyde products of lipid peroxidation and to allyl alcohol (Bilinski et al 1989) and also to cis 3-nonen-1-ol, a metabolic precursor of nonenal (Biliński et al 2005). It should be mentioned that increased formation of ROS has been found to follow exposure of another yeast, Candida albicans, to allyl alcohol (Lemar et al 2005).…”

“…The effects of treatment of S. cerevisiae cells with allyl alcohol resemble those of acrolein exposure including disruption of cellular kinetics and of redox balance (Wills and Phelps 1978), peroxidation of cellular lipids, depletion of intracellular stores of glutathione, and in consequence, decrease in cell viability (Biliński et al 2005;Trotter et al 2006).…”