Lipid particles of the yeast Saccharomyces cerevisiae are storage compartments for triacylglycerols (TAG) and steryl esters (STE). Four gene products, namely the TAG synthases Dga1p and Lro1p, and the STE synthases Are1p and Are2p contribute to storage lipid synthesis. A yeast strain lacking the four respective genes is devoid of lipid particles thus providing a valuable tool to study the physiological role of storage lipids and lipid particles. Using a dga1lro1are1are2 quadruple mutant transformed with plasmids bearing inducible DGA1, LRO1, or ARE2 we demonstrate that TAG synthesis contributes more efficiently to lipid particle proliferation than synthesis of STE. Moreover, we show that proteins typically located to lipid particles in wild type such as Erg1p, Erg6p, Erg7p, and Ayr1p are refined to microsomal fractions of the dga1lro1are1are2 quadruple mutant. This result confirms the close relationship between lipid particles and endoplasmic reticulum. Most interestingly, the amount of the squalene epoxidase Erg1p, which is dually located in lipid particles and endoplasmic reticulum of wild type, is decreased in the quadruple mutant, whereas amounts of other lipid particle proteins tested were not reduced. This decrease is not caused by down-regulation of ERG1 transcription but by the low stability of Erg1p in the quadruple mutant. Because a similar effect was also observed in are1are2 mutants this finding can be mainly attributed to the lack of STE. The quadruple mutant, however, was more sensitive to terbinafine, an inhibitor of Erg1p, than the are1are2 strain suggesting that the presence of TAG and/or intact lipid particles has an additional protective effect. In a strain lacking the two STE synthases, Are1p and Are2p, incorporation of ergosterol into the plasma membrane was reduced, although the total cellular amount of free ergosterol was higher in the mutant than in wild type. Thus, an esterification/deacylation mechanism appears to contribute to the supply of ergosterol to the plasma membrane.

• PMID: 15155725
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Journal Article | Research Support, Non-U.S. Gov't

Authors

Sorger D, Athenstaedt K, Hrastnik C, Daum G

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