Oxidative stress promotes specific protein damage in Saccharomyces cerevisiae

Abstract

We have analyzed the proteins that are oxidatively damaged when Saccharomyces cerevisiae cells are exposed to stressing conditions. Carbonyl groups generated by hydrogen peroxide or menadione on proteins of aerobically respiring cells were detected by Western blotting, purified, and identified. Mitochondrial proteins such as E2 subunits of both pyruvate dehydrogenase and alpha-ketoglutarate dehydrogenase, aconitase, heat-shock protein 60, and the cytosolic fatty acid synthase (alpha subunit) and glyceraldehyde-3-phosphate dehydrogenase were the major targets. In addition we also report the in vivo modification of lipoamide present in the above-mentioned E2 subunits under the stressing conditions tested and that this also occurs with the homologous enzymes present in Escherichia coli cells that were used for comparative analysis. Under fermentative conditions, the main protein targets in S. cerevisiae cells treated with hydrogen peroxide or menadione were pyruvate decarboxylase, enolase, fatty acid synthase, and glyceraldehyde-3-phosphate dehydrogenase. Under the stress conditions tested, fermenting cells exhibit a lower viability than aerobically respiring cells and, consistently, increased peroxide generation as well as higher content of protein carbonyls and lipid peroxides. Our results strongly suggest that the oxidative stress in prokaryotic and eukaryotic cells shares common features.

Changes in disulfide bond content of proteins in a yeast strain lacking major sources of NADPH

“…In particular, all of the glycolytic and cytosolic metabolic enzymes listed in Table 3, with the exception of Lys9p, were identified in both types of studies [24,46,47]. In other functional categories, Sod1p and cyclophilin Cpr1p were similarly identified as carbonylated in the Δidp2Δzwf1 mutant shifted to oleate medium, and in wild-type glucose-grown hydrogen peroxide-challenged cells [24].…”

“…(These effects were much less pronounced with a shift of the mutant strain to acetate medium.) We used two-dimensional electrophoresis to examine carbonylated proteins in the mutant strain shifted to oleate in the absence or presence of glutathione and DTT, with the goal of comparing results with our data on disulfide bond-containing proteins, as well as with results of several reported studies of carbonylation in yeast cells grown on glucose with exogenous oxidants [24,46,47].…”

Rice ASR1 Protein with Reactive Oxygen Species Scavenging and Chaperone-like Activities Enhances Acquired Tolerance to Abiotic Stresses in Saccharomyces cerevisiae

“…Pgk1p was abundantly expressed in cells grown in glucose, and transcription was increased by heat shock (Piper et al, 1986) and regulated by the transcription factors Rab1p, Abf1p, and Reb1p (Packham et al, 1996). In particular, enolase and Ssb protein are major targets of protein damage in WT yeast cells exposed to oxidative stress (Cabiscol et al, 2000;Reverter-Branchat et al, 2004). Downregulation of glycolytic enzymes by irreversible protein oxidation impairs glucose utilization and this effect is correlated with enhanced gluconeogenic and energy storage pathways, and causes an imbalance in proteostasis, which plays a role in protein translocation, folding, and assembly (Reverter-Branchat et al, 2004).…”

Does age influence loss of heterozygosity?

“…Protein oxidation is known to increase with age, and can lead to protein dysfunction (Stadtman, 1992). In addition oxidative modification does not affect the proteome uniformly, in that some proteins are more prevalent targets (Cabiscol et al, 2000). In studies of budding yeast, carbonylated proteins were visualized in single cells and seen to accumulate with increased replicative age (Aguilaniu et al, 2003).…”