当肌醇磷酸化神经酰胺合成酶Aur1失活时,酿酒酵母依赖高尔基体和液泡之间的囊泡运输。

Eukaryotic Cell Pub Date : 2015-12-01 Epub Date: 2015-10-02 DOI:10.1128/EC.00117-15
Natalia S Voynova, Carole Roubaty, Hector M Vazquez, Shamroop K Mallela, Christer S Ejsing, Andreas Conzelmann
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引用次数: 11

摘要

肌醇磷酸化神经酰胺(IPC)及其甘露糖基化衍生物是酵母中唯一的复杂鞘脂。它们的合成可以被特异性抑制IPC合成酶Aur1的aureobasidin A (AbA)减少。据报道,AbA通过降低IPC水平,导致内质网(ER)应激,胞质钙增加,活性氧产生和线粒体损伤导致细胞凋亡。我们发现,当Aur1通过转录下调逐渐耗尽时,神经酰胺的积累成为细胞存活的主要障碍。在这种情况下,碱性神经酰胺酶YPC1的过度表达可以拯救细胞。我们建立了羟基化C26脂肪酸作为神经酰胺水解的可靠标志。这种水解仅在YPC1过表达时发生。相反,当Aur1被AbA急性抑制时,YPC1的过表达没有任何有益作用。一项高通量遗传筛选显示,当Aur1被抑制时,无论其抑制方式如何,高尔基体、核内体和液泡之间的囊泡介导运输对存活至关重要。此外,当细胞受到AbA的急性应激时,液泡酸化是必不可少的,而向液泡摄取奎宁表明,AbA激活了液泡酸化。抗氧化剂n -乙酰半胱氨酸对AbA作用下的细胞生长没有促进作用,表明AbA诱导的活性氧自由基在其毒性中起次要作用。AbA强烈诱导细胞壁完整性通路,但渗透支持不能提高野生型细胞在AbA作用下的生存能力。总之,这些数据支持并完善了aba介导的细胞死亡的现有模型,并将液泡蛋白转运和酸化作为抗病性的新关键因素。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Saccharomyces cerevisiae Is Dependent on Vesicular Traffic between the Golgi Apparatus and the Vacuole When Inositolphosphorylceramide Synthase Aur1 Is Inactivated.

Saccharomyces cerevisiae Is Dependent on Vesicular Traffic between the Golgi Apparatus and the Vacuole When Inositolphosphorylceramide Synthase Aur1 Is Inactivated.

Saccharomyces cerevisiae Is Dependent on Vesicular Traffic between the Golgi Apparatus and the Vacuole When Inositolphosphorylceramide Synthase Aur1 Is Inactivated.

Saccharomyces cerevisiae Is Dependent on Vesicular Traffic between the Golgi Apparatus and the Vacuole When Inositolphosphorylceramide Synthase Aur1 Is Inactivated.

Inositolphosphorylceramide (IPC) and its mannosylated derivatives are the only complex sphingolipids of yeast. Their synthesis can be reduced by aureobasidin A (AbA), which specifically inhibits the IPC synthase Aur1. AbA reportedly, by diminishing IPC levels, causes endoplasmic reticulum (ER) stress, an increase in cytosolic calcium, reactive oxygen production, and mitochondrial damage leading to apoptosis. We found that when Aur1 is gradually depleted by transcriptional downregulation, the accumulation of ceramides becomes a major hindrance to cell survival. Overexpression of the alkaline ceramidase YPC1 rescues cells under this condition. We established hydroxylated C26 fatty acids as a reliable hallmark of ceramide hydrolysis. Such hydrolysis occurs only when YPC1 is overexpressed. In contrast, overexpression of YPC1 has no beneficial effect when Aur1 is acutely repressed by AbA. A high-throughput genetic screen revealed that vesicle-mediated transport between Golgi apparatus, endosomes, and vacuole becomes crucial for survival when Aur1 is repressed, irrespective of the mode of repression. In addition, vacuolar acidification becomes essential when cells are acutely stressed by AbA, and quinacrine uptake into vacuoles shows that AbA activates vacuolar acidification. The antioxidant N-acetylcysteine does not improve cell growth on AbA, indicating that reactive oxygen radicals induced by AbA play a minor role in its toxicity. AbA strongly induces the cell wall integrity pathway, but osmotic support does not improve the viability of wild-type cells on AbA. Altogether, the data support and refine current models of AbA-mediated cell death and add vacuolar protein transport and acidification as novel critical elements of stress resistance.

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来源期刊
Eukaryotic Cell
Eukaryotic Cell 生物-微生物学
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期刊介绍: Eukaryotic Cell (EC) focuses on eukaryotic microbiology and presents reports of basic research on simple eukaryotic microorganisms, such as yeasts, fungi, algae, protozoa, and social amoebae. The journal also covers viruses of these organisms and their organelles and their interactions with other living systems, where the focus is on the eukaryotic cell. Topics include: - Basic biology - Molecular and cellular biology - Mechanisms, and control, of developmental pathways - Structure and form inherent in basic biological processes - Cellular architecture - Metabolic physiology - Comparative genomics, biochemistry, and evolution - Population dynamics - Ecology
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