title: Low ergosterol content in yeast adh1 mutant enhances chitin maldistribution and sensitivity to paraquat-induced oxidative stress.
abstract: Alcohol dehydrogenases catalyse the reversible oxidation of alcohols to aldehydes or ketones, with concomitant reduction of NAD(+) or NADP(+) . Adh1p is responsible for the reduction of acetaldehyde to ethanol, while Adh2p catalyses the reverse reaction, the oxidation of ethanol to acetaldehyde. Lack of Adh1p shifts the cellular redox balance towards excess NADH/NADPH and acetaldehyde, while absence of Adh2p does the opposite. Yeast mutant adh1Δ had a slow growth rate, whereas adh2Δ grew like the isogenic wild-type (WT) during prediauxic shift fermentative metabolism. After 48 h WT and mutants reached the same number of viable cells. When exponentially growing (LOG) cells were exposed to calcofluor white, only mutant adh1Δ displayed an irregular deposition of chitin. Quantitative analyses of both LOG and stationary-phase cells showed that adh1Δ mutant contained significantly less ergosterol than cells of WT and adh2Δ mutant, whereas the erg3Δ mutant contained extremely low ergosterol pools. Both adh1Δ and adh2Δ mutants showed higher-than-WT resistance to heat shock and to H(2) O(2) but had WT resistance when exposed to ultraviolet (UV) light and the DNA cross-linking agent diepoxyoctane, indicating normal DNA repair capacity. Mutant adh1Δ was specifically sensitive to acetaldehyde and to membrane peroxidizing paraquat. Our results link the pleiotropic phenotype of adh1Δ mutants to low pools of ergosterol and to reductive stress, and introduce the two new phenotypes, resistance to heat shock and to H(2) O(2) , for the adh2Δ mutant, most probably related to increased ROS production in mitochondria, which leads to the induction of oxidative stress protection.
authors.affiliation: Laboratório de Biologia de Fungos, Centro de Biotecnologia e Genética, Universidade Estadual de Santa Cruz, Rod. Ilhéus Itabuna km 16, Salobrinho, Ilhéus, Bahia, Brazil.
authors.forename: G
authors.initials: G
authors.lastname: Marisco
authors.forename: S T
authors.initials: ST
authors.lastname: Saito
authors.forename: I S
authors.initials: IS
authors.lastname: Ganda
authors.forename: M
authors.initials: M
authors.lastname: Brendel
authors.forename: C
authors.initials: C
authors.lastname: Pungartnik
mesh_terms: D000426:Alcohol Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D001557:Benzenesulfonates / Q000737:chemistry; D002686:Chitin / Q000378:metabolism*; D004875:Ergosterol / Q000378:metabolism*; D018869:Heat-Shock Response / Q000502:physiology; D008856:Microscopy, Fluorescence; D009154:Mutation; D018384:Oxidative Stress / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D010269:Paraquat / Q000494:pharmacology*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D001557:Benzenesulfonates; D029701:Saccharomyces cerevisiae Proteins; D002686:Chitin; C007061:C.I. Fluorescent Brightening Agent 28; C516801:ADH1 protein, S cerevisiae; D000426:Alcohol Dehydrogenase; D010269:Paraquat; D004875:Ergosterol