O'Doherty P J, Lyons V, Higgins V J, Rogers P J, Bailey T D, Wu M J
School of Science and Health, University of Western Sydney , Penrith, New South Wales , Australia.
Free Radic Res. 2013 Dec;47(12):1054-65. doi: 10.3109/10715762.2013.849344. Epub 2013 Oct 21.
Eukaryotic microorganisms are constantly challenged by reactive oxygen species derived endogenously or encountered in their environment. Such adversity is particularly applied to Saccharomyces cerevisiae under harsh industrial conditions. One of the major oxidants to challenge S. cerevisiae is linoleic acid hydroperoxide (LoaOOH). This study, which used genome-wide microarray analysis in conjunction with deletion mutant screening, uncovered the molecular pathways of S. cerevisiae that were altered by an arresting concentration of LoaOOH (75 μM). The oxidative stress response, iron homeostasis, detoxification through PDR transport and direct lipid β-oxidation were evident through the induction of the genes encoding for peroxiredoxins (GPX2, TSA2), the NADPH:oxidoreductase (OYE3), iron uptake (FIT2, ARN2, FET3), PDR transporters (PDR5, PDR15, SNQ2) and β-oxidation machinery (FAA2, POX1). Further, we discovered that Gpx3p, the dual redox sensor and peroxidase, is required for protection against LoaOOH, indicated by the sensitivity of gpx3Δ to a mild dose of LoaOOH (37.5 μM). Deletion of GPX3 conferred a greater sensitivity to LoaOOH than the loss of its signalling partner YAP1. Deletion of either of the iron homeostasis regulators AFT1 or AFT2 also resulted in sensitivity to LoaOOH. These novel findings for Gpx3p, Aft1p and Aft2p point to their distinct roles in response to the lipid peroxide. Finally, the expression of 89 previously uncharacterised genes was significantly altered against LoaOOH, which will contribute to their eventual annotation.
真核微生物不断受到内源性产生或在其环境中遇到的活性氧的挑战。在恶劣的工业条件下,这种逆境尤其适用于酿酒酵母。挑战酿酒酵母的主要氧化剂之一是亚油酸氢过氧化物(LoaOOH)。本研究结合全基因组微阵列分析和缺失突变体筛选,揭示了酿酒酵母在LoaOOH(75μM)的抑制浓度下发生改变的分子途径。通过诱导编码过氧化物酶(GPX2、TSA2)、NADPH:氧化还原酶(OYE3)、铁摄取(FIT2、ARN2、FET3)、PDR转运蛋白(PDR5、PDR15、SNQ2)和β-氧化机制(FAA2、POX1)的基因,氧化应激反应、铁稳态、通过PDR转运进行解毒以及直接脂质β-氧化明显可见。此外,我们发现双氧化还原传感器和过氧化物酶Gpx3p是抵御LoaOOH所必需的,gpx3Δ对低剂量LoaOOH(37.5μM)的敏感性表明了这一点。缺失GPX3比缺失其信号伴侣YAP1对LoaOOH更敏感。缺失铁稳态调节因子AFT1或AFT2也会导致对LoaOOH敏感。这些关于Gpx3p、Aft1p和Aft2p的新发现表明它们在应对脂质过氧化物方面具有独特作用。最后,89个先前未表征基因的表达在LoaOOH作用下发生了显著改变,这将有助于它们最终的注释。