Systematic analysis of yeast strains with possible defects in lipid metabolism

Daum, Günther; Tuller, Gabriele; Nemec, Tamara; Hrastnik, Cladia; Balliano, Gianni; Cattel, Luigi; Milla, Paola; Rocco, Flavio; Conzelmann, Aadreas; Vionnet, Christine; Kelly, Diane; Kelly, Steven L.; Schweizer, Eckhard; Schüller, Hans‐Joachim; Hojad, Ursula; Greiner, Eva; Finger, Kerin

doi:10.1002/(sici)1097-0061(199905)15:7<601::aid-yea390>3.0.co;2-n

Cited by 160 publications

(118 citation statements)

References 20 publications

(13 reference statements)

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“…S3B). Although it is commonly believed that PC dominate the yeast glycerophospholipidome (23), our analysis of wild-type strains BY4742, BY4743, NY13 and CTY182 consistently identified PI as a major lipid class constituting 17% to 30% of the lipidome.…”

Section: Resultsmentioning

confidence: 60%

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Global analysis of the yeast lipidome by quantitative shotgun mass spectrometry

Ejsing

Sampaio

Surendranath

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

967

1,064

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Although the transcriptome, proteome, and interactome of several eukaryotic model organisms have been described in detail, lipidomes remain relatively uncharacterized. Using Saccharomyces cerevisiae as an example, we demonstrate that automated shotgun lipidomics analysis enabled lipidome-wide absolute quantification of individual molecular lipid species by streamlined processing of a single sample of only 2 million yeast cells. By comparative lipidomics, we achieved the absolute quantification of 250 molecular lipid species covering 21 major lipid classes. This analysis provided Ϸ95% coverage of the yeast lipidome achieved with 125-fold improvement in sensitivity compared with previous approaches. Comparative lipidomics demonstrated that growth temperature and defects in lipid biosynthesis induce ripple effects throughout the molecular composition of the yeast lipidome. This work serves as a resource for molecular characterization of eukaryotic lipidomes, and establishes shotgun lipidomics as a powerful platform for complementing biochemical studies and other systems-level approaches.fatty acid elongation ͉ S. cerevisiae ͉ shotgun lipidomics T he lipidome of eukaryotic cells consists of hundreds to thousands of individual lipid species that constitute membranes, store metabolic energy and function as bioactive molecules (1-3). Despite the extensive characterization of proteins, their association into complexes and activities (4-6), it is still difficult to assess how perturbations within the lipid metabolic network affect the full lipidome of cells. This work shows that lipidome-wide quantification of individual molecular lipid species (molecules with defined chemical structure) by absolute quantification (expressed in mol or mol%) provides a new approach to relate lipidomics and functional genomics studies.The yeast Saccharomyces cerevisiae serves as a prime model organism for studying the molecular organization and regulatory circuitry of eukaryotic lipidomes (7-9). It uses a relatively simple and conserved network of lipid metabolic pathways (Fig. 1) that synthesize a few hundred molecular lipid species constituting its full lipidome (3). The lipidome diversity is primarily determined by the fatty acid synthase (10), the ⌬-9 desaturase (11) and the fatty acid elongation complex (12) that produce only saturated or mono-unsaturated fatty acids having 10 to 26 carbon atoms for the biosynthesis of glycerolipids, glycerophospholipids, and sphingolipids. Importantly, several metabolic conversions interlink sphingolipid, glycerophospholipid, and glycerolipid metabolism such that any perturbation within the metabolic network is prone to induce lipidome-wide ripple effects. Remarkably, numerous genes involved in lipid metabolism and trafficking can be mutated or deleted without apparent physiological consequences (Fig. 1).Despite remarkable methodological advances, lipidomics seldom complements functional genomics efforts owing to three major factors. First, analysis of glycerophospholipids and sphingolipids requires ...

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Section: Resultsmentioning

confidence: 60%

“…The most abundant lipid species was ergosterol comprising 12.0 mol% of the lipidome (equivalent to 481 pmol/0.2 OD units), followed by 8. (16,23).…”

Section: Resultsmentioning

confidence: 99%

Global analysis of the yeast lipidome by quantitative shotgun mass spectrometry

Ejsing

Sampaio

Surendranath

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

967

1,064

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“…Three polypeptides encoded by YDL109c, YGL144c, and YDR444w contain lipase-active site motifs (www.proteome. com/), and a mutant deleted of YDL109c exhibits increased TAG levels (17). None of these open reading frames, however, was demonstrated by enzymatic analysis of the gene product to encode a TAG lipase.…”

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confidence: 99%

YMR313c/TGL3 Encodes a Novel Triacylglycerol Lipase Located in Lipid Particles of Saccharomyces cerevisiae

Athenstaedt

Daum

2003

Journal of Biological Chemistry

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191

162

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Previous work from our laboratory (Athenstaedt, K., Zweytick, D., Jandrositz, A., Kohlwein, S. D., and Daum, G. (1999) J. Bacteriol. 181, 6441-6448) showed that the gene product of YMR313c (named Tgl3p) is a component of yeast lipid particles, and deletion of this gene led to an increase in the cellular level of triacylglycerols (TAG). These observations suggested that TGL3 may encode a TAG lipase of Saccharomyces cerevisiae. Here we demonstrate by cell fractionation and by microscopic inspection of a strain bearing a Tgl3p-GFP hybrid that this polypeptide is highly enriched in the lipid particle fraction but virtually absent from other organelles. The entire TAG lipase activity of lipid particles is attributed to Tgl3p, because the activity in this organelle is completely absent in a ⌬tgl3 deletion mutant, whereas it is significantly enhanced in a strain overexpressing Tgl3p. A His 6 -tagged Tgl3p hybrid purified close to homogeneity from a yeast strain overexpressing this fusion protein exhibited high TAG lipase activity. Most importantly, experiments in vivo using the fatty acid synthesis inhibitor cerulenin demonstrated that deletion of TGL3 resulted in a decreased mobilization of TAG from lipid particles. The amino acid sequence deduced from the open reading frame YMR313c contains the consensus sequence motif GXSXG typical for lipolytic enzymes. Otherwise, Tgl3p has no significant sequence homology to other lipases identified so far. In summary, our data identified Tgl3p as a novel yeast TAG lipase at the molecular level and by function in vivo and in vitro.

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“…It is interesting to note that according to previously reported research 7 Saccharomyces cerevisiae cannot produce 18: 2. The strain used in our study does not only de- viate from other Saccharomyces cerevisiae strains by showing both Flo1 and NewFlo behaviours, but also in regard to its lipid metabolism.…”

Section: Lipid Turnover In Flo1 Phenotype Behaviourmentioning

confidence: 81%

Lipid Turnover During Inverse Flocculation in Saccharomyces cerevisiae UOFS Y-2330

Strauss

Kock

Viljoen

et al. 2004

Journal of the Institute of Brewing

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In this study we uncovered that Saccharomyces cerevisiae UOFS Y-2330 does not only demonstrate inverse flocculation, but is also characterised by two different lipid turnover patterns. During Flo1 phenotype flocculation, this yeast showed two neutral lipid accumulating stages (i.e. at 8 h and from 12 h). This is probably triggered by flocculation, which can be regarded as a survival mechanism where cells accumulate predominantly neutral lipids as a reserve energy source -a similar mechanism is probably operative when cells enter stationary growth. Contrary to Flo1 behaviour, this strain in NewFlo phenotype mode demonstrates only a single lipid accumulation phase i.e. when cells enter stationary growth, which coincides with increase in flocculation. In addition, an increase in phospholipids was experienced during active growth in both flocculation behaviours i.e. Flo1 and NewFlo probably as a result of active membrane production.

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Systematic analysis of yeast strains with possible defects in lipid metabolism

Cited by 160 publications

References 20 publications

Global analysis of the yeast lipidome by quantitative shotgun mass spectrometry

Global analysis of the yeast lipidome by quantitative shotgun mass spectrometry

YMR313c/TGL3 Encodes a Novel Triacylglycerol Lipase Located in Lipid Particles of Saccharomyces cerevisiae

Lipid Turnover During Inverse Flocculation in Saccharomyces cerevisiae UOFS Y-2330

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