Overexpression of the White Opaque Switching Master Regulator Wor1 Alters Lipid Metabolism and Mitochondrial Function in Candida albicans
Abstract
:1. Introduction
2. Materials and Methods
2.1. Strains and Growth Conditions
2.2. Molecular Biology Procedures and Plasmid Constructions
2.3. RNA Isolation and RT-qPCR Analysis
2.4. Mitochondrial Function Assays
2.5. Protein and Immunoblot Methods
2.6. Analysis of Lipid Content
3. Results
3.1. Lipid Content Is Altered in WOR1OE Strain
3.2. WOR1 Overexpression Enhances the Utilization of Phospholipids and Increases the Number and Size of Peroxisomes
3.3. WOR1 Overexpression Alters the Susceptibility to Certain Membrane Disturbing Compounds
3.4. Mitochondrial Function Is Compromised by WOR1 Overexpression
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Strain Name (Original) | Background Strain and Genotype | Reference |
---|---|---|
SC5314 | Clinical isolate | [32] |
CAI4 | ura3Δ::imm434/ura3Δ::imm434 iro1/iro1::imm434 | [33] |
CAI4-pNRUe (REP40) | [CAI4] ADH1/adh1::tTATETPR-myc-URA3 | [34] |
CAI4-RFP | [CAI4] ADH1/adh1::TDH3PRtTATETPR-dTOM2-URA3 | [31] |
CAI4-WOR1OE | [CAI4] ADH1/adh1::TDH3PRtTATETPR-WOR1-myc-URA3 | [31] |
SU6 | [CAI4-pNRUe] ARD1/ard1::TDH3PR-GFP-PXP2M SAT1 | This study |
SU7 | [CAI4-WOR1OE] ARD1/ard1::TDH3PR-GFP-PXP2M SAT1 | This study |
hog1 ura3 (HI7) | [CAI4] hog1::hisG/hog1::hisG | [35] |
hog1-pNRUe | [hog1 ura3] ADH1/adh1::tTATETPR-myc-URA3 | Román, submitted |
hog1-WOR1OE | [hog1 ura3] ADH1/adh1::TDH3PRtTATeTPR-WOR1-myc-URA3 | Román, submitted |
Primer | Sequence (5′–3′) | Reference |
---|---|---|
o-ACTQTup | TGGTGGTTCTATCTTGGCTTCA | [38] |
o-ACTQTlw | ATCCACATTTGTTGGAAAGTAGA | [38] |
o-ERG11up-QT | TTACCTCATTATTGGAGACGTGATG | This study |
o-ERG11lw-QT | CACCACGTTCTCTTCTCAGTTTAATT | This study |
o1-PXP2 | AATGCGGCCGCATGGCTATGCTTACTAAATCTATACATGATG | This study |
o2-PXP2 | ACATGGATTTGGTCCCATTGATGGTG | This study |
o3-PXP2 | CACCATCAATGGGACCAAATCCATGT | This study |
o4-PXP2 | GAAGCAGCTGCTAAATTATCAAGATAATTAGATCTAAT | This study |
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Hidalgo-Vico, S.; Casas, J.; García, C.; Lillo, M.P.; Alonso-Monge, R.; Román, E.; Pla, J. Overexpression of the White Opaque Switching Master Regulator Wor1 Alters Lipid Metabolism and Mitochondrial Function in Candida albicans. J. Fungi 2022, 8, 1028. https://doi.org/10.3390/jof8101028
Hidalgo-Vico S, Casas J, García C, Lillo MP, Alonso-Monge R, Román E, Pla J. Overexpression of the White Opaque Switching Master Regulator Wor1 Alters Lipid Metabolism and Mitochondrial Function in Candida albicans. Journal of Fungi. 2022; 8(10):1028. https://doi.org/10.3390/jof8101028
Chicago/Turabian StyleHidalgo-Vico, Susana, Josefina Casas, Carolina García, M. Pilar Lillo, Rebeca Alonso-Monge, Elvira Román, and Jesús Pla. 2022. "Overexpression of the White Opaque Switching Master Regulator Wor1 Alters Lipid Metabolism and Mitochondrial Function in Candida albicans" Journal of Fungi 8, no. 10: 1028. https://doi.org/10.3390/jof8101028