Cited by 113Antifungal resistance and susceptibilityFungal and yeast genetics researchFungal Plant Pathogen Control

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Facultative Sterol Uptake in an Ergosterol-Deficient Clinical Isolate of Candida glabrata Harboring a Missense Mutation in ERG11 and Exhibiting Cross-Resistance to Azoles and Amphotericin B

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Hull, Claire M., Parker, Josie E., Bader, Oliver, Weig, Michael, Gross, Uwe, Warrilow, Andrew G. S., Kelly, Diane E., Kelly, Steven L.·2012·Antimicrobial Agents and Chemotherapy

DOI 10.1128/aac.06253-11PMC PMC3421581

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Abstract

We identified a clinical isolate of Candida glabrata (CG156) exhibiting flocculent growth and cross-resistance to fluconazole (FLC), voriconazole (VRC), and amphotericin B (AMB), with MICs of >256, >256, and 32 μg ml(-1), respectively. Sterol analysis using gas chromatography-mass spectrometry (GC-MS) revealed that CG156 was a sterol 14α-demethylase (Erg11p) mutant, wherein 14α-methylated intermediates (lanosterol was >80% of the total) were the only detectable sterols. ERG11 sequencing indicated that CG156 harbored a single-amino-acid substitution (G315D) which nullified the function of native Erg11p. In heterologous expression studies using a doxycycline-regulatable Saccharomyces cerevisiae erg11 strain, wild-type C. glabrata Erg11p fully complemented the function of S. cerevisiae sterol 14α-demethylase, restoring growth and ergosterol synthesis in recombinant yeast; mutated CG156 Erg11p did not. CG156 was culturable using sterol-free, glucose-containing yeast minimal medium ((glc)YM). However, when grown on sterol-supplemented (glc)YM (with ergosta 7,22-dienol, ergosterol, cholestanol, cholesterol, Δ(7)-cholestenol, or desmosterol), CG156 cultures exhibited shorter lag phases, reached higher cell densities, and showed alterations in cellular sterol composition. Unlike comparator isolates (harboring wild-type ERG11) that became less sensitive to FLC and VRC when cultured on sterol-supplemented (glc)YM, facultative sterol uptake by CG156 did not affect its azole-resistant phenotype. Conversely, CG156 grown using (glc)YM with ergosterol (or with ergosta 7,22-dienol) showed increased sensitivity to AMB; CG156 grown using (glc)YM with cholesterol (or with cholestanol) became more resistant (MICs of 2 and >64 μg AMB ml(-1), respectively). Our results provide insights into the consequences of sterol uptake and metabolism on growth and antifungal resistance in C. glabrata.

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Authors: Hull, C. M., Parker, J. E., Bader, O., Weig, M., Gross, U., Warrilow, A. G. S., Kelly, D. E., & Kelly, S. L.
Affiliations: Swansea University (GB)
Universitätsmedizin Göttingen (DE)
ORCID: Parker, Josie E.
Bader, Oliver
Weig, Michael
Gross, Uwe
Warrilow, Andrew G. S.
Kelly, Diane E.
Kelly, Steven L.
Venue: Antimicrobial Agents and Chemotherapy
Publisher: American Society for Microbiology
Published: 2012-08
Volume: 56
Issue: 8
Pages: 4223-4232
ISSN: 0066-4804 (ISSN-L)
1098-6596
DOI: 10.1128/aac.06253-11
PMC: PMC3421581
OpenAlex: W2128071392
Cited by: 113
References: 42
Citations by year: 2026: 4 · 2025: 9 · 2024: 8 · 2023: 7 · 2022: 13 · 2021: 11 · 2020: 12 · 2019: 6 · 2018: 2 · 2017: 14 · 2016: 7 · 2015: 4 · 2014: 10 · 2013: 5 · 2012: 1
FWCI: 4.42
Type: journal article
Language: en
Open access: green
europepmc.org
Licence: journals.asm.org/non-commercial-tdm-license (tdm)
Locations: Antimicrobial Agents and Chemotherapy (publishedVersion)
Europe PMC (PubMed Central) (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
University of Groningen research database (University of Groningen / Centre for Information Technology) (submittedVersion)
Funders: Engineering and Physical Sciences Research Council
Topics: Health Sciences › Medicine › Infectious Diseases › Antifungal resistance and susceptibility
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
Life Sciences › Agricultural and Biological Sciences › Plant Science › Fungal Plant Pathogen Control
Keywords: Candida glabrata, ERG11, sterol 14α-demethylase, missense mutations, azole resistance, amphotericin B resistance, cross-resistance, flocculation, sterol composition, lanosterol, cholesterol, cholestanol, desmosterol, antifungal resistance, clinical isolates
MeSH terms: Amphotericin B, Antifungal Agents, Azoles, Base Sequence, Biological Transport, Ergosterol, Fungal Proteins, Humans, Microbial Sensitivity Tests, Pyrimidines, Saccharomyces cerevisiae, Sterols, Triazoles, Fluconazole, Sequence Analysis, DNA, Mutation, Missense, Drug Resistance, Multiple, Fungal, Candida glabrata, Sterol 14-Demethylase, Voriconazole

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Facultative Sterol Uptake in an Ergosterol-Deficient Clinical Isolate of Candida glabrata Harboring a Missense Mutation in ERG11 and Exhibiting Cross-Resistance to Azoles and Amphotericin B

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