Mechanistic role of ergosterol in membrane rigidity and cycloheximide resistance in Saccharomyces cerevisiae

Abe, Fumiyoshi; Hiraki, Toshiki

doi:10.1016/j.bbamem.2008.12.002

Cited by 151 publications

(119 citation statements)

References 48 publications

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“…2B). However, measurement of membrane order with the probe trimethylammonium-DPH (TMA-DPH), which labels exclusively the plasma membrane surface due to its TMA moiety (52), indicates that there is a decrease of its order in erg6⌬ cells, and no alteration in scs7⌬ cells, when compared with WT cells (2). A different behavior of DPH and TMA-DPH is better explained by a distinct change in the domain structure of intracellular and plasma membranes.…”

Section: Discussionmentioning

confidence: 99%

Gel Domains in the Plasma Membrane of Saccharomyces cerevisiae

Aresta‐Branco

Cordeiro

Marinho

et al. 2011

Journal of Biological Chemistry

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The plasma membrane of Saccharomyces cerevisiae was studied using the probes trans-parinaric acid and diphenylhexatriene. Diphenylhexatriene anisotropy is a good reporter of global membrane order. The fluorescence lifetimes of transparinaric acid are particularly sensitive to the presence and nature of ordered domains, but thus far they have not been measured in yeast cells. A long lifetime typical of the gel phase (>30 ns) was found in wild-type (WT) cells from two different genetic backgrounds, at 24 and 30°C, providing the first direct evidence for the presence of gel domains in living cells. To understand their nature and location, the study of WT cells was extended to spheroplasts, the isolated plasma membrane, and liposomes from total lipid and plasma membrane lipid extracts (with or without ergosterol extraction by cyclodextrin). It is concluded that the plasma membrane is mostly constituted by ordered domains and that the gel domains found in living cells are predominantly at the plasma membrane and are formed by lipids. To understand their composition, strains with mutations in sphingolipid and ergosterol metabolism and in the glycosylphosphatidylinositol anchor remodeling pathway were also studied. The results strongly indicate that the gel domains are not ergosterol-enriched lipid rafts; they are mainly composed of sphingolipids, possibly inositol phosphorylceramide, and contain glycosylphosphatidylinositol-anchored proteins, suggesting an important role in membrane traffic and signaling, and interactions with the cell wall. The abundance of the sphingolipid-enriched gel domains was inversely related to the cellular membrane system global order, suggesting their involvement in the regulation of membrane properties.

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Section: Discussionmentioning

confidence: 99%

Gel Domains in the Plasma Membrane of Saccharomyces cerevisiae

Aresta‐Branco

Cordeiro

Marinho

et al. 2011

Journal of Biological Chemistry

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“…The properties of the membrane of living yeast cells were analyzed as described previously (27). BY4743 yeast cells (OD 600 of 0.1) were inoculated into synthetic medium containing the solvent (1% [vol/vol] ethanol), 0.0015% (vol/vol) NP-40, and 40 g of GlcCer/ml purified from soybean and incubated for 17 h without shaking.…”

Section: Methodsmentioning

confidence: 99%

Glucosylceramide Contained in Koji Mold-Cultured Cereal Confers Membrane and Flavor Modification and Stress Tolerance to Saccharomyces cerevisiae during Coculture Fermentation

Sawada

Sato

Hamajima

et al. 2015

Appl Environ Microbiol

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fIn nature, different microorganisms create communities through their physiochemical and metabolic interactions. Many fermenting microbes, such as yeasts, lactic acid bacteria, and acetic acid bacteria, secrete acidic substances and grow faster at acidic pH values. However, on the surface of cereals, the pH is neutral to alkaline. Therefore, in order to grow on cereals, microbes must adapt to the alkaline environment at the initial stage of colonization; such adaptations are also crucial for industrial fermentation. Here, we show that the yeast Saccharomyces cerevisiae, which is incapable of synthesizing glucosylceramide (GlcCer), adapted to alkaline conditions after exposure to GlcCer from koji cereal cultured with Aspergillus kawachii. We also show that various species of GlcCer derived from different plants and fungi similarly conferred alkali tolerance to yeast. Although exogenous ceramide also enhanced the alkali tolerance of yeast, no discernible degradation of GlcCer to ceramide was observed in the yeast culture, suggesting that exogenous GlcCer itself exerted the activity. Exogenous GlcCer also increased ethanol tolerance and modified the flavor profile of the yeast cells by altering the membrane properties. These results indicate that GlcCer from A. kawachii modifies the physiology of the yeast S. cerevisiae and demonstrate a new mechanism for cooperation between microbes in food fermentation. In nature, microbial communities are formed through metabolic and physiochemical interactions among different microorganisms (1). The pH of cereals that have ripened and dropped from the husk to the ground is neutral to alkaline (2-4). Fermentation microbes, including Saccharomyces cerevisiae, Lactobacillus lactis, and Acetobacter aceti, efficiently utilize carbohydrates in the cereals to produce organic acids (1). As a result, the microbial communities that colonize such carbohydrate-rich cereals, as well as those found in fermented foods, reduce the pH to 4.0 to 6.0. After these microbes establish an acidic pH, they can dominate the environment because they can grow faster at acidic pH values than other microbes. Therefore, these microbes must first adapt to the alkaline environment at the initial stage of colonization on cereal; such adaptation mechanisms are also crucial for industrial fermentation.Traditional alcoholic beverages are produced via the fermentation of cereals by the yeast S. cerevisiae, which can efficiently catabolize glucose to produce ethanol. However, because S. cerevisiae is incapable of hydrolyzing starch to glucose, additional biological catalysts are often added to the fermentation reaction. For example, malt catalyzes starch hydrolysis in wheat-and barleyderived beverages such as whiskey and beer. In East and Southeast Asia, various cereals fermented by fungi are widely used as starchhydrolyzing catalysts for the production of rice-derived alcoholic beverages. These include Japanese sake (5, 6) and shochu, Korean Makgeolli, and Chinese Huangjiu, as well as various other fermented foods ...

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“…9A). To verify the plasma membrane localization of CgFtr1-GFP and CgFet3-GFP, wt and Cgvps34⌬ cells were labeled with TMA-DPH (trimethylamino-diphenyl-1,3,5-hexatriene), which marks solely the plasma membrane (34). As shown in Fig.…”

Section: Transcriptional Response To Changes In the Environmental Iromentioning

confidence: 99%

The Phosphoinositide 3-Kinase Regulates Retrograde Trafficking of the Iron Permease CgFtr1 and Iron Homeostasis in Candida glabrata

Sharma

Purushotham

Kaur

2016

Journal of Biological Chemistry

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Edited by James SiedowThe phosphoinositide 3-kinase (PI3K), which phosphorylates phosphatidylinositol and produces PI3P, has been implicated in protein trafficking, intracellular survival, and virulence in the pathogenic yeast Candida glabrata. Here, we demonstrate PI3-kinase (CgVps34) to be essential for maintenance of cellular iron homeostasis. We examine how CgVps34 regulates the fundamental process of iron acquisition, and underscore its function in vesicular trafficking as a central determinant. RNA sequencing analysis revealed iron homeostasis genes to be differentially expressed upon CgVps34 disruption. Consistently, the Cgvps34⌬ mutant displayed growth attenuation in low-and high-iron media, increased intracellular iron content, elevated mitochondrial aconitase activity, impaired biofilm formation, and extenuated mouse organ colonization potential. Furthermore, we demonstrate for the first time that C. glabrata cells respond to iron limitation by expressing the iron permease CgFtr1 primarily on the cell membrane, and to iron excess via internalization of the plasma membrane-localized CgFtr1 to the vacuole. Our data show that CgVps34 is essential for the latter process. We also report that macrophage-internalized C. glabrata cells express CgFtr1 on the cell membrane indicative of an iron-restricted macrophage internal milieu, and Cgvps34⌬ cells display better survival in iron-enriched medium-cultured macrophages. Overall, our data reveal the centrality of PI3K signaling in iron metabolism and host colonization.

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Mechanistic role of ergosterol in membrane rigidity and cycloheximide resistance in Saccharomyces cerevisiae

Cited by 151 publications

References 48 publications

Gel Domains in the Plasma Membrane of Saccharomyces cerevisiae

Gel Domains in the Plasma Membrane of Saccharomyces cerevisiae

Glucosylceramide Contained in Koji Mold-Cultured Cereal Confers Membrane and Flavor Modification and Stress Tolerance to Saccharomyces cerevisiae during Coculture Fermentation

The Phosphoinositide 3-Kinase Regulates Retrograde Trafficking of the Iron Permease CgFtr1 and Iron Homeostasis in Candida glabrata

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