Abstract
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 ergΔ mutants. DRM fractionation revealed that the association between Pma1-GFP and DRM was weakened in erg6Δ 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 ergA 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.
| Original language | English |
|---|---|
| Pages (from-to) | 830-841 |
| Number of pages | 12 |
| Journal | Microbiology |
| Volume | 154 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - Mar 1 2008 |
| Externally published | Yes |
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All Science Journal Classification (ASJC) codes
- Microbiology
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Multiple functions of ergosterol in the fission yeast Schizosaccharomyces pombe. / Iwaki, Tomoko; Iefuji, Haruyuki; Hiraga, Yoshikazu; Hosomi, Akira; Morita, Tomotake; Giga-Hama, Yuko; Takegawa, Kaoru.
In: Microbiology, Vol. 154, No. 3, 01.03.2008, p. 830-841.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Multiple functions of ergosterol in the fission yeast Schizosaccharomyces pombe
AU - Iwaki, Tomoko
AU - Iefuji, Haruyuki
AU - Hiraga, Yoshikazu
AU - Hosomi, Akira
AU - Morita, Tomotake
AU - Giga-Hama, Yuko
AU - Takegawa, Kaoru
PY - 2008/3/1
Y1 - 2008/3/1
N2 - 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 ergΔ mutants. DRM fractionation revealed that the association between Pma1-GFP and DRM was weakened in erg6Δ 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 ergA 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.
AB - 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 ergΔ mutants. DRM fractionation revealed that the association between Pma1-GFP and DRM was weakened in erg6Δ 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 ergA 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.
UR - http://www.scopus.com/inward/record.url?scp=42949144293&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=42949144293&partnerID=8YFLogxK
U2 - 10.1099/mic.0.2007/011155-0
DO - 10.1099/mic.0.2007/011155-0
M3 - Article
C2 - 18310029
AN - SCOPUS:42949144293
VL - 154
SP - 830
EP - 841
JO - Microbiology
JF - Microbiology
SN - 1350-0872
IS - 3
ER -