Lactate Like Fluconazole Reduces Ergosterol Content in the Plasma Membrane and Synergistically Kills Candida albicans
Abstract
:1. Introduction
2. Results
2.1. Lactate Affects the Resistance of C. albicans to Fluconazole Depending on Ergosterol—The First Observations
2.2. Cdr1 Transporter Activity and CDR1 Gene Expression Are More Efficiently Upregulated in the Presence Lactate Compared with Glucose
2.3. Lactate Accelerates Delocalization of Cdr1 Transporter from PM into Vacuoles
2.4. Lactate Inhibits Ergosterol Synthesis in C. albicans Cells
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Strains and Growth Conditions
4.3. MIC50 Determination
4.4. Microscopic Studies
4.5. Western Blot
4.6. Real-Time PCR
4.7. Efflux Activity of Cdr1 Transporter
4.8. Phenotype Assay
4.9. PM Isolation and Sterol Analysis
4.10. Statistical Analyses
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Strain C. albicans | Medium | Fluconazole | Brefeldin A | Fluphenazine |
---|---|---|---|---|
WT | YPD | 2 | 16 | 125 |
YPL | 0.5 | 32 | >250 | |
cdr1Δ | YPD | 0.25 | 4 | 125 |
YPL | 0.063 | 4 | 250 | |
cdr2Δ | YPD | 2 | 16 | 125 |
YPL | 0.5 | 32 | >250 | |
cdr1Δcdr2Δ | YPD | 0.25 | 4 | 62.5 |
YPL | 0.063 | 4 | 62.5 |
Glucose | Lactate | |||
---|---|---|---|---|
8 h | 14 h | 8 h | 14 h | |
Ergosterol | 39.1 ± 2.3 | 65.2 ± 1.1 *** | 22.9 ± 0.3 ** | 17.8 ± 4.8 *** |
Lanosterol | 8.5 ± 0.2 | 5.4 ± 0.6 ** | 5.9 ± 0.5 ** | 5.1 ± 0.8 ** |
4-methylfecosterol | ND | ND | 3.9 ± 0.5 | 3.9 ± 0.2 |
Ergosta-5,7-dienol | ND | ND | 1.1 ± 0.1 | 0.8 ± 0.2 |
4,4-dimethylcholesta-8,24-dien-3b-ol | ND | ND | 1.3 ± 0.3 | 1.1 ± 0.1 |
Total sterols | 47.6 ± 2.5 | 70.6 ± 1.7 *** | 35.1 ± 1.7 | 28.7 ± 6.1 ** |
Strain | Genotype | Reference |
---|---|---|
CAF2-1 | ura3Δ::imm434/URA3 | [37] |
DSY448 | ura3∆::imm434/ura3∆::imm434 cdr1∆::hisG/cdr1∆::hisG-URA3-hisG | [38] |
DSY653 | ura3∆::imm434/ura3∆::imm434 cdr2∆::hisG/cdr2∆::hisG-URA3-hisG | [39] |
DSY654 | ura3∆::imm434/ura3∆::imm434 cdr1∆::hisG/cdr1∆::hisG cdr2∆::hisG/cdr2∆::hisG-URA3-hisG | [39] |
ASCa1 | ura3Δ::imm434/ura3Δ::imm434 CDR1/CDR1-yEGFP-URA3 | [40] |
KS028 | ura3Δ::imm434/ura3Δ::imm434 erg11Δ::SAT1-FLIP/erg11Δ::FRT | [21] |
Primer | Sequence 5′–3′ |
---|---|
RDN18F | AGAAACGGCTACCACATCCAA |
RDN18R | GGGCCCTGTATCGTTATTTATTGT |
ERG11F | TTTGGTGGTGGTAGACATA |
ERG11R | GAACTATAATCAGGGTCAGG |
ERG3F | CCATCATGAATCATGACAGTCC |
ERG3R | TGCTTCTCATGCTTTCCATC |
CDR1F | TTTAGCCAGAACTTTCACTCATGATT |
CDR1R | TATTTATTTCTTCATGTTCATATGGATTGA |
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Suchodolski, J.; Muraszko, J.; Bernat, P.; Krasowska, A. Lactate Like Fluconazole Reduces Ergosterol Content in the Plasma Membrane and Synergistically Kills Candida albicans. Int. J. Mol. Sci. 2021, 22, 5219. https://doi.org/10.3390/ijms22105219
Suchodolski J, Muraszko J, Bernat P, Krasowska A. Lactate Like Fluconazole Reduces Ergosterol Content in the Plasma Membrane and Synergistically Kills Candida albicans. International Journal of Molecular Sciences. 2021; 22(10):5219. https://doi.org/10.3390/ijms22105219
Chicago/Turabian StyleSuchodolski, Jakub, Jakub Muraszko, Przemysław Bernat, and Anna Krasowska. 2021. "Lactate Like Fluconazole Reduces Ergosterol Content in the Plasma Membrane and Synergistically Kills Candida albicans" International Journal of Molecular Sciences 22, no. 10: 5219. https://doi.org/10.3390/ijms22105219