Departamento de Biologia, Centro de Biologia Molecular e Ambiental, Universidade do Minho, Braga, Portugal.
Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.
Oxid Med Cell Longev. 2020 Apr 2;2020:7095078. doi: 10.1155/2020/7095078. eCollection 2020.
The yeast undergoes a mitochondrial-dependent regulated cell death (RCD) exhibiting typical markers of mammalian apoptosis. We have previously shown that ceramide production contributes to RCD induced by acetic acid and is involved in mitochondrial outer membrane permeabilization and cytochrome release, especially through hydrolysis of complex sphingolipids catalyzed by Isc1p. Recently, we also showed that Sch9p regulates the translocation of Isc1p from the endoplasmic reticulum into mitochondria, perturbing sphingolipid balance and determining cell fate. In this study, we addressed the role of other signaling proteins in acetic acid-induced RCD. We found that single deletion of or , as shown for and , leads to an increase in cell survival in response to acetic acid and that Pkh1/2p-dependent phosphorylation of Ypk1p and Sch9p increases under these conditions. These results indicate that Pkh1p regulates acetic acid-induced RCD through Ypk1p and Sch9p. In addition, our results suggest that Pkh1p-Ypk1p is necessary for resistance to acetic acid-induced RCD. Moreover, double deletion of and has a drastic effect on cell survival associated with increased ROS accumulation and release of cytochrome , which is counteracted by overexpression of the PKA pathway negative regulator . Overall, our results suggest that Pkh1p-Ypk1p and Pkh1p-Sch9p pathways contribute to RCD induced by acetic acid.
酵母经历依赖于线粒体的调控性细胞死亡（RCD），表现出典型的哺乳动物细胞凋亡特征。我们之前已经表明，神经酰胺的产生有助于乙酸诱导的 RCD，并参与线粒体外膜通透性和细胞色素 c 的释放，特别是通过 Isc1p 催化的复杂鞘脂的水解。最近，我们还表明 Sch9p 调节 Isc1p 从内质网向线粒体的易位，扰乱鞘脂平衡并决定细胞命运。在这项研究中，我们研究了其他信号蛋白在乙酸诱导的 RCD 中的作用。我们发现，如 和 所示，缺失 或 会导致细胞对乙酸的存活能力增加，并且在这些条件下 Pkh1/2p 依赖性磷酸化 Ypk1p 和 Sch9p 增加。这些结果表明 Pkh1p 通过 Ypk1p 和 Sch9p 调节乙酸诱导的 RCD。此外，我们的结果表明 Pkh1p-Ypk1p 是对乙酸诱导的 RCD 的 抗性所必需的。此外，缺失 和 会对细胞存活产生严重影响，与 ROS 积累增加和细胞色素 c 的释放有关，而过表达 PKA 途径的负调节剂 可以抵消这种影响。总体而言，我们的结果表明 Pkh1p-Ypk1p 和 Pkh1p-Sch9p 途径有助于乙酸诱导的 RCD。
