The Sensitivity of Yeast Mutants to Oleic Acid Implicates the Peroxisome and Other Processes in Membrane Function

Lockshon, Daniel; Surface, Lauren E.; Kerr, Emily O.; Kaeberlein, Matt; Kennedy, Brian K.

doi:10.1534/genetics.106.064428

Cited by 99 publications

(118 citation statements)

References 87 publications

(85 reference statements)

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“…1A). These findings are in agreement with those from a genome-wide screen performed with yeast cells of a closely related genetic background (15). In addition, wild type cells carrying Gat1-GFP were able to grow in the presence of oleic acid confirming that the fusion protein is functional (12).…”

Section: Cells Lacking Gat1p Do Not Accumulate Lipid Particlessupporting

confidence: 89%

“…Cells lacking Gat1p, but not Gat2p, are sensitive to oleic acid (this study and Ref. 15). Staining of oleate-grown gat1⌬ cells with Nile Red followed by fluorescence microscopy revealed a failure in LP accumulation contrasting to wild type and gat2⌬ strains.…”

Section: Discussionsupporting

confidence: 56%

“…Excess availability of fatty acids, like oleate, provides such an experimental condition (5,14). Interestingly, in a recent screening of the haploid yeast deletion collection, cells lacking Gat1p (but not Gat2p) were identified to be sensitive to oleate (15). Here, we demonstrate that Gat1p is indeed a major player in the cellular response to oleic acid, markedly contrasting with Gat2p.…”

mentioning

confidence: 64%

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Controlling Lipid Fluxes at Glycerol-3-phosphate Acyltransferase Step in Yeast

Marr

Foglia

Terebiznik

et al. 2012

Journal of Biological Chemistry

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Background: Excess oleate induces triacylglycerol synthesis and proliferation of lipid particles (LP) in yeast. Results: Oleate-induced LP formation is dependent on the glycerol-3-phosphate acyltransferase (GPAT) isoform Gat1p, which physically interacts with LPs. Conclusion: Formation of oleate-induced LPs depends on active Gat1p (but not Gat2p). Significance: Fatty acid surplus is controlled at the GPAT step with unique contributions from different isoforms.

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Section: Cells Lacking Gat1p Do Not Accumulate Lipid Particlessupporting

confidence: 89%

Section: Discussionsupporting

confidence: 56%

mentioning

confidence: 64%

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Controlling Lipid Fluxes at Glycerol-3-phosphate Acyltransferase Step in Yeast

Marr

Foglia

Terebiznik

et al. 2012

Journal of Biological Chemistry

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“…Indeed, when Gpd1p localization was altered, cell growth was dramatically compromised when cells were challenged with both oleic acid and osmotic stress. In this case, the cell is faced with the need to increase glycerol concentration through increased glycolysis but also must metabolize the oleic acid, which can be toxic otherwise (48). Both processes produce and consume NADH and are controlled in part by the NAD ϩ /NADH ratio.…”

Section: Discussionmentioning

confidence: 99%

“…Gpd1p Is Not Required for Peroxisomal ␤-Oxidation-Because we and others have not detected differences between gpd1⌬ and wild-type strains in large scale screens of growth in the presence of fatty acid (48,49), we used more sensitive measures to determine whether Gpd1p is required for peroxisome biogenesis or peroxisomal ␤-oxidation. Liquid growth assays were done using a Bioscreen to compare the growth rates of wild-type, pex3⌬, and gpd1⌬ cells in glucose medium and oleic acid medium (Fig.…”

Section: Gpd1p Localization To Peroxisomes Is Regulated By Phosphorylmentioning

confidence: 99%

Dynamic Changes in the Subcellular Distribution of Gpd1p in Response to Cell Stress

Jung

Marelli

Rachubinski

et al. 2010

Journal of Biological Chemistry

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Gpd1p is a cytosolic NAD؉ -dependent glycerol 3-phosphate dehydrogenase that also localizes to peroxisomes and plays an essential role in the cellular response to osmotic stress and a role in redox balance. Here, we show that Gpd1p is directed to peroxisomes by virtue of an N-terminal type 2 peroxisomal targeting signal (PTS2) in a Pex7p-dependent manner. Significantly, localization of Gpd1p to peroxisomes is dependent on the metabolic status of cells and the phosphorylation of aminoacyl residues adjacent to the targeting signal. Exposure of cells to osmotic stress induces changes in the subcellular distribution of Gpd1p to the cytosol and nucleus. This behavior is similar to Pnc1p, which is coordinately expressed with Gpd1p, and under conditions of cell stress changes its subcellular distribution from peroxisomes to the nucleus where it mediates chromatin silencing. Although peroxisomes are necessary for the ␤-oxidation of fatty acids in yeast, the localization of Gpd1p to peroxisomes is not. Rather, shifts in the distribution of Gpd1p to different cellular compartments in response to changing cellular status suggests a role for Gpd1p in the spatial regulation of redox potential, a process critical to cell survival, especially under the complex stress conditions expected to occur in the wild.Glycerol 3-phosphate dehydrogenase (Gpd1p) is one of two NAD ϩ -dependent glycerol 3-phosphate dehydrogenases in yeast (1, 2). It is classically defined as a cytosolic enzyme that catalyzes the conversion of dihydroxyacetone phosphate (DHAP) 2 and NADH to glycerol 3-phosphate (Glycerol 3-phosphate) and NAD ϩ (2). In unstressed cells, this reaction prevents the accumulation of DHAP (3), which can otherwise be transformed into methyl glyoxylate (MG) (3, 4), a toxic compound that interacts with proteins (5-7). This reaction also allows for the reoxidation of NADH to NAD ϩ , which can serve as a buffer for cytosolic redox balance, compensating for cellular reactions that produce NADH (2). Moreover, glycerol 3-phosphate is a key metabolite for the synthesis of glyceride lipids and phospholipids, as well as for the formation of glycerol (8 -12).Under hyperosmotic stress, Saccharomyces cerevisiae, as well as other yeasts, accumulates glycerol as a major solute (13,14). This increased production of glycerol is caused mainly by an enhanced activity of Gpd1p, and accordingly, Gpd1p is essential for growth under osmotic stress (2, 15). In addition, GPD1 expression is altered by a wide variety of stresses, including heat, cold, and oxidative stress (16 -18), suggesting its regulation is controlled by stress. Genome-wide monitoring of transcript changes in yeast under various stress conditions also showed that GPD1 belongs to a large group of common stressresponse genes (19, 20); however, its regulation is complex and appears to be controlled by multiple signaling pathways (21). This likely reflects the multiple metabolic changes that yeast cells undergo in response to different stresses (19,22,23) and the function of Gpd1p at the int...

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A metabolic engineering strategy for producing free fatty acids by the Yarrowia lipolytica yeast based on impairment of glycerol metabolism

Yuzbasheva

Mostova

Андреева

et al. 2017

Biotech & Bioengineering

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In recent years, bio-based production of free fatty acids from renewable resources has attracted attention for their potential as precursors for the production of biofuels and biochemicals. In this study, the oleaginous yeast Yarrowia lipolytica was engineered to produce free fatty acids by eliminating glycerol metabolism. Free fatty acid production was monitored under lipogenic conditions with glycerol as a limiting factor. Firstly, the strain W29 (Δgpd1), which is deficient in glycerol synthesis, was obtained. However, W29 (Δgpd1) showed decreased biomass accumulation and glucose consumption in lipogenic medium containing a limiting supply of glycerol. Analysis of substrate utilization from a mixture of glucose and glycerol by the parental strain W29 revealed that glycerol was metabolized first and glucose utilization was suppressed. Thus, the Δgpd1Δgut2 double mutant, which is deficient also in glycerol catabolism, was constructed. In this genetic background, growth was repressed by glycerol. Oleate toxicity was observed in the Δgpd1Δgut2Δpex10 triple mutant strain which is deficient additionally in peroxisome biogenesis. Consequently, two consecutive rounds of selection of spontaneous mutants were performed. A mutant released from growth repression by glycerol was able to produce 136.8 mg L of free fatty acids in a test tube, whereas the wild type accumulated only 30.2 mg L . Next, an isolated oleate-resistant strain produced 382.8 mg L of free fatty acids. Finely, acyl-CoA carboxylase gene (ACC1) over-expression resulted to production of 1436.7 mg L of free fatty acids. The addition of dodecane promoted free fatty acid secretion and enhanced the level of free fatty acids up to 2033.8 mg L during test tube cultivation.

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The Sensitivity of Yeast Mutants to Oleic Acid Implicates the Peroxisome and Other Processes in Membrane Function

Cited by 99 publications

References 87 publications

Controlling Lipid Fluxes at Glycerol-3-phosphate Acyltransferase Step in Yeast

Controlling Lipid Fluxes at Glycerol-3-phosphate Acyltransferase Step in Yeast

Dynamic Changes in the Subcellular Distribution of Gpd1p in Response to Cell Stress

A metabolic engineering strategy for producing free fatty acids by the Yarrowia lipolytica yeast based on impairment of glycerol metabolism

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