School of Science and Health, Western Sydney University, Locked Bag 1797, Penrith NSW 2751, Australia.
Australian Proteome Analysis Facility (APAF), Macquarie University, Sydney NSW 2109 Australia.
FEMS Yeast Res. 2017 May 1;17(3). doi: 10.1093/femsyr/fox022.
Yeast AP-1 transcription factor (Yap1p) and the enigmatic oxidoreductases Oye2p and Oye3p are involved in counteracting lipid oxidants and their unsaturated breakdown products. In order to uncover the response to linoleic acid hydroperoxide (LoaOOH) and the roles of Oye2p, Oye3p and Yap1p, we carried out proteomic analysis of the homozygous deletion mutants oye3Δ, oye2Δ and yap1Δ alongside the diploid parent strain BY4743. The findings demonstrate that deletion of YAP1 narrowed the response to LoaOOH, as the number of proteins differentially expressed in yap1Δ was 70% of that observed in BY4743. The role of Yap1p in regulating the major yeast peroxiredoxin Tsa1p was demonstrated by the decreased expression of Tsa1p in yap1Δ. The levels of Ahp1p and Hsp31p, previously shown to be regulated by Yap1p, were increased in LoaOOH-treated yap1Δ, indicating their expression is also regulated by another transcription factor(s). Relative to BY4743, protein expression differed in oye3Δ and oye2Δ under LoaOOH, underscored by superoxide dismutase (Sod1p), multiple heat shock proteins (Hsp60p, Ssa1p, and Sse1p), the flavodoxin-like protein Pst2p and the actin stabiliser tropomyosin (Tpm1p). Proteins associated with glycolysis were increased in all strains following treatment with LoaOOH. Together, the dataset reveals, for the first time, the yeast proteomic response to LoaOOH, highlighting the significance of carbohydrate metabolism, as well as distinction between the roles of Oye3p, Oye2p and Yap1p.
酵母 AP-1 转录因子（Yap1p）和神秘的氧化还原酶 Oye2p 和 Oye3p 参与对抗脂质氧化剂及其不饱和分解产物。为了揭示对亚油酸氢过氧化物（LoaOOH）的反应以及 Oye2p、Oye3p 和 yap1p 的作用，我们对同源缺失突变体 oye3Δ、oye2Δ 和 yap1Δ 以及二倍体亲本菌株 BY4743 进行了蛋白质组分析。研究结果表明，YAP1 的缺失缩小了对 LoaOOH 的反应范围，因为 yap1Δ 中差异表达的蛋白质数量是 BY4743 中观察到的蛋白质数量的 70%。Yap1p 在调节主要酵母过氧化物酶 Tsa1p 中的作用通过 yap1Δ 中 Tsa1p 的表达减少得到证明。Ahp1p 和 Hsp31p 的水平先前被证明受 yap1p 调节，在 LoaOOH 处理的 yap1Δ 中增加，表明它们的表达也受另一个转录因子（s）调节。与 BY4743 相比，LoaOOH 处理下 oye3Δ 和 oye2Δ 的蛋白质表达不同，突出了超氧化物歧化酶（Sod1p）、多种热休克蛋白（Hsp60p、Ssa1p 和 Sse1p）、黄嘌呤氧化酶样蛋白 Pst2p 和肌动蛋白稳定剂原肌球蛋白（Tpm1p）。LoaOOH 处理后，所有菌株的糖酵解相关蛋白均增加。总的来说，该数据集首次揭示了酵母对 LoaOOH 的蛋白质组反应，强调了碳水化合物代谢的重要性，以及 Oye3p、Oye2p 和 yap1p 作用的区别。
