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Sterol synthesis and viability oferg11 (cytochrome P450 lanosterol demethylase) mutations inSaccharomyces cerevisiae andCandida albicans

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Lipids

Abstract

The identification of the precise structural features of yeast sterol molecules required for the essential “sparking” function has been a controversial area of research. Recent cloning and gene disruption studies inSaccharomyces cerevisiae have shown that C-24 methylation (ERG6), C-5 desaturation (ERG3) and Δ8-Δ7 isomerization (ERG2) are not required, while C-14 demethylation (ERG11) and C-14 reduction (ERG24) are each required for aerobic viability. Earlier observations had indicated that C-14 demethylase deficient strains could be restored to aerobic growth by suppressor mutations that caused a deficiency in C-5 desaturase. These strains were reported to synthesize some ergosterol, indicating that they contained leaky mutations in bothERG11 andERG3, thereby making it imposssible to determine whether the removal of the C-14 methyl group was required for aerobic viability. The availability of theERG11 andERG3 genes has been used in this study to construct strains that contain null mutants in bothERG11 andERG3. Results show that these double disruption strains are viable and that spontaneously arising suppressors of theERG11 disruption areerg3 mutants. Theerg11 mutants ofS. cerevisiae are compared to similar mutants ofCandida albicans that are viable in the absence of theerg3 lesion.

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Abbreviations

BSTFA:

N,O-bis(trimethylsilyl)trifluoroacetamide

GC:

gas chromatography

MS:

mass spectroscopy

PCR:

polymerase chain reaction

sld :

suppressor of lanosterol demethylation

TMCS:

trimethylchlorosilane

Tween 80:

polyoxyethylenesorbitan monooleate

YMG:

galactose minimal medium

YMM:

yeast minimal medium

YPD:

yeast extract, peptone, dextrose

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Authors and Affiliations

  1. Department of Biology, Indiana University-Purdue University at Indianapolis, 723 W. Michigan St., 46202-5132, Indianapolis, Indiana

    M. Bard, N. D. Lees & L. Cofrin

  2. Department of Molecular Genetics, University of Cincinnati College of Medicine, 45267, Cincinnati, Ohio

    T. Turi & J. C. Loper

  3. Department of Environmental Health, University of Cincinnati College of Medicine, 45267, Cincinnati, Ohio

    D. Craft & J. C. Loper

  4. Marion Merrell Dow Pharmaceutical Inc., 45215, Cincinnati, Ohio

    R. Barbuch & C. Koegel

Authors
  1. M. Bard
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  2. N. D. Lees
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  3. T. Turi
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  4. D. Craft
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  5. L. Cofrin
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  6. R. Barbuch
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  7. C. Koegel
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  8. J. C. Loper
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Bard, M., Lees, N.D., Turi, T. et al. Sterol synthesis and viability oferg11 (cytochrome P450 lanosterol demethylase) mutations inSaccharomyces cerevisiae andCandida albicans . Lipids 28, 963–967 (1993). https://doi.org/10.1007/BF02537115

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  • DOI: https://doi.org/10.1007/BF02537115

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