Cloning, disruption and sequence of the gene encoding yeast C-5 sterol desaturase

Arthington, Beth A.; Bennett, Linda G.; Skatrud, Paul L.; Guynn, Cynthia J.; Barbuch, Robert J.; UIbright, Corinne E.; Bard, Martin

doi:10.1016/0378-1119(91)90535-j

Cited by 139 publications

(123 citation statements)

References 17 publications

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“…149,216 Subsequently, the encoding yeast ERG3 gene was cloned and disruption of the gene resulted in a strain that retained viability under aerobic conditions. 5 The resolution of these contradictory conclusions lies in consideration of other markers in the strains used. 244 ERG3 disruptions were done in a heme-competent strain, while feeding experiments were accomplished in a heme-deficient background.…”

Section: Sterol Biosynthesis In Yeast: Farnesyl Pyrophosphate To Ergomentioning

confidence: 99%

Biochemistry, cell biology and molecular biology of lipids ofSaccharomyces cerevisiae

Daum

Lees

Bard

et al. 1998

Yeast

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The yeast Saccharomyces cerevisiae is a powerful experimental system to study biochemical, cell biological and molecular biological aspects of lipid synthesis. Most but not all genes encoding enzymes involved in fatty acid, phospholipid, sterol or sphingolipid biosynthesis of this unicellular eukaryote have been cloned, and many gene products have been functionally characterized. Less information is available about genes and gene products governing the transport of lipids between organelles and within membranes, turnover and degradation of complex lipids, regulation of lipid biosynthesis, and linkage of lipid metabolism to other cellular processes. Here we summarize current knowledge about lipid biosynthetic pathways in S. cerevisiae and describe the characteristic features of the gene products involved. We focus on recent discoveries in these fields and address questions on the regulation of lipid synthesis, subcellular localization of lipid biosynthetic steps, cross‐talk between organelles during lipid synthesis and subcellular distribution of lipids. Finally, we discuss distinct functions of certain key lipids and their possible roles in cellular processes. © 1998 John Wiley & Sons, Ltd.

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Section: Sterol Biosynthesis In Yeast: Farnesyl Pyrophosphate To Ergomentioning

confidence: 99%

Biochemistry, cell biology and molecular biology of lipids ofSaccharomyces cerevisiae

Daum

Lees

Bard

et al. 1998

Yeast

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380

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“…Mutations in erg2 and erg3 both result in a 6-fold increase in ERG3 expression. The erg2 mutation in S. cerevisiae is non-auxotrophic for sterol and results in an accumulation of sterol intermediates such as ergosta-5,8,22-313-ol, and ergosta-8-en-313-ol [15] and the erg3 mutation leads to an accumulation of ergosta-7,22-dien-313-ol [3]. Similarly, a mutation in the HMG1 gene encoding the major isoform of HMG-CoA reductase [16] results in a 4-fold increase in ERG3 expression.…”

Section: A Block In the Ergosterol Biosynthetic Pathway Up-regulatmentioning

confidence: 99%

“…E-mail: iujk 100@indyvax.iupui.edu kingdoms. We have shown previously by gene-disruption that ERG3 is non-essential for the aerobic viability of Saccharomyces cerevisiae [3]. Haploid yeast cells harboring a null allele of ERG3 accumulate the sterol intermediate episterol (ergosta-7,22-dien-3~-ol) which can functionally substitute for ergosterol in the membrane.…”

Section: Introductionmentioning

confidence: 99%

Positive and negative regulation of a sterol biosynthetic gene (ERG3) in the post‐squalene portion of the yeast ergosterol pathway

et al. 1996

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Regulation of sterol biosynthesis in the terminal portion of the pathway represents an efficient mechanism by which the cell can control the production of sterol without disturbing the production of other essential mevaionate pathway products. We demonstrate that mutations affecting early and late steps in sterol homeostasis modulate the expression of the ERG3 gene: a late step in sterol biosynthesis in yeast. Expression of ERG3 is increased in response to a mutation in the major isoform of HMG CoA reductase which catalyzes the rate-limiting step of sterol biosynthesis. Likewise, mutations in non-auxotrophic ergosterol biosynthetic genes downstream of squalene production (erg2, erg3, erg4, ergS, and ergO') result in an up-regulation of ERG3 expression. Deletion analysis of the ERG3 promoter identified two upstream activation sequences: UAS1, which when deleted reduces ERG3 gene expression 3-4-fuld but maintains sterol regulation and UAS2, which when deleted further reduces ERG3 expression and abolishes sterol regulation. The recent isolation of two yeast genes responsible for the esterification of intracellular sterol (ARE1 and ARE2) has enabled us to directly analyze the relationship between sterol esterification and de novo biosynthesis. Our results demonstrate that the absence of sterol esterification leads to a decrease in total intracellular sterol and ERG3 is a target of this negative regulation.

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“…A single ERG3 gene is found in Sacch. cerevisiae, Candida albicans and Candida glabrata, while Aspergillus fumigatus has three ERG3 homologues (Arthington et al, 1991;Geber et al, 1995;Miyazaki et al, 1999; Alcazar-Fuoli et al, 2006). While ergosterol was not detected in the double disruption mutant erg31Derg32D, it was present in the single mutants.…”

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Multiple functions of ergosterol in the fission yeast Schizosaccharomyces pombe

et al. 2008

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Sterols are a major class of membrane lipids in eukaryotes. In Schizosaccharomyces pombe, sterol 24-C-methyltransferase (Erg6p), C-8 sterol isomerase (Erg2p), C-5 sterol desaturase (Erg31p, Erg32p), C-22 sterol desaturase (Erg5p) and C-24 (28) sterol reductase (Sts1p/ Erg4p) have been predicted, but not yet determined, to catalyse a sequence of reactions from zymosterol to ergosterol. Disruption mutants of these genes were unable to synthesize ergosterol, and most were tolerant to the polyene drugs amphotericin B and nystatin. Disruption of erg31 + or erg32 + did not cause ergosterol deficiency or tolerance to polyene drugs, indicating that the two C-5 sterol desaturases have overlapping functions. GFP-tagged DRM (detergent-resistant membrane)-associated protein Pma1p localized to the plasma membrane in ergD mutants. DRM fractionation revealed that the association between Pma1-GFP and DRM was weakened in erg6D but not in other erg mutants. Several GFP-tagged plasma membrane proteins were tested, and an amino acid permease homologue, SPBC359.03c, was found to mislocalize to intracellular punctate structures in the ergD mutants. These results indicate that these proteins are responsible for ergosterol biosynthesis in fission yeast, similar to the situation in Saccharomyces cerevisiae. Furthermore, in fission yeast, ergosterol is important for plasma membrane structure and function and for localization of plasma membrane proteins. INTRODUCTIONSterols are essential structural and regulatory components of eukaryotic cell membranes. Mammals, plants and fungi produce similar sterols, which differ in the number and location of double bonds and methyl side chains. Ergosterol is the end product of the sterol biosynthetic pathway and is the major sterol in yeasts (Fig. 1a). Like cholesterol in mammalian cells, it is responsible for membrane fluidity and permeability (Parks et al., 1995).Ergosterol synthesis and metabolism have been well defined in the budding yeast Saccharomyces cerevisiae (Daum et al., 1998). Multiple genetic and biochemical studies have culminated in the virtually complete elucidation of the pathway leading to ergosterol (Lees et al., 1995;Parks et al., 1995). The enzymic reactions have been largely defined by identification of erg mutants defective in ergosterol biosynthesis, and by their complementation based on sterol auxotrophy or altered sterol composition. Most genes involved in the early part of the pathway to lanosterol are essential for growth, because yeasts require sterols and no sterol molecule is synthesized up to this point. In contrast, mutations in the steps from zymosterol to ergosterol (Fig. 1b) are not essential for growth because the intermediates produced can partially substitute for ergosterol. The latter part of the ergosterol biosynthetic pathway is not linear in the sense of consecutive reactions, because the enzymes converting lanosterol to ergosterol do not show a strict substrate preference. Thus, null mutants impaired in the steps from zymosterol to ergosterol accumulate char...

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Cloning, disruption and sequence of the gene encoding yeast C-5 sterol desaturase

Cited by 139 publications

References 17 publications

Biochemistry, cell biology and molecular biology of lipids ofSaccharomyces cerevisiae

Biochemistry, cell biology and molecular biology of lipids ofSaccharomyces cerevisiae

Positive and negative regulation of a sterol biosynthetic gene (ERG3) in the post‐squalene portion of the yeast ergosterol pathway

Multiple functions of ergosterol in the fission yeast Schizosaccharomyces pombe

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