The Glycosylphosphatidylinositol-Anchored Superoxide Dismutase of Scedosporium apiospermum Protects the Conidia from Oxidative Stress
Abstract
:1. Introduction
2. Materials and Methods
2.1. Strain and Culture Conditions
2.2. Genomic DNA Extraction
2.3. Disruption of the SODD Gene
2.4. Southern Blot Analysis
2.5. Susceptibility Studies
2.6. Phagocytosis Assays
2.7. Transmission Electron Microscopy
2.8. Statistical Analysis
3. Results
3.1. Generation of a SODD Deficient Mutant
3.2. Sensitivity to Temperature and Cell Wall Stressing Chemicals
3.3. Growth under Chemically-Induced Oxidative Stress
3.4. Susceptibility to Antifungals
3.5. Interactions with Phagocytes
3.6. Morphological Features of the Conidia and Ultrastructure of Their Cell Wall
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Primer Name and Use | Sequence 5′→3′ | Tm (°C) | Size of the Amplicon (bp) |
---|---|---|---|
PCR amplification of the 5′ flanking region of SODD | |||
P1-SODD | ATTCATAGACTCAATAATTAGAACTCGACT | 64 | 925 |
P2-SODD | TCGTGAATCTTTTACCAGATCGGAAGCAATAAATGTAATTTATCTCTTTCAATCCCAAGC | 64 | |
PCR amplification of the hygromycin B resistance gene | |||
P3-SODD | GCTTGGGATTGAAAGAGATAAATTACATTTATTGCTTCCGATCTGGTAAAAGATTCACGA | 68 | 2633 |
P4-SODD | AATTGATTCTTGTCGATCATTAATTTGGTCATCAGAGCAGATTGTACTGAGAGTGCACCA | 68 | |
PCR amplification of the 5′ flanking region of SODD | |||
P5-SODD | TGGTGCACTCTCAGTACAATCTGCTCTGATGACCAAATTAATGATCGACAAGAATCAATT | 64 | 901 |
P6-SODD | GACGTTGTATATATATCCTGGAAGAATCTT | 64 | |
Fusion of the amplicons | |||
P7-SODD | GAAACGCCCGACTAGTTAAATC | 64 | 4448 |
P8-SODD | CTGCAAATGCCAAATTCCAA | 64 |
Antifungal | Parent Strain (KU70Δ Mutant) | KU70Δ/SODDΔ Double Mutant | p-Value | ||
---|---|---|---|---|---|
MIC Mean (µg/mL) | SD | MIC Mean (µg/mL) | SD | ||
Isavuconazole | >32 | ND | 0.142 | 0.093 | <0.05 |
Itraconazole | 0.552 | 0.187 | 0.064 | 0.000 | <0.05 |
Posaconazole | 0.476 | 0.053 | 0.131 | 0.035 | <0.05 |
Voriconazole | 0.251 | 0.146 | 0.018 | 0.004 | <0.05 |
Fluconazole | >256 | ND | >256 | ND | NS |
Amphotericin B | 5 | 2 | 3.9 | 1.597 | NS |
Caspofungin | >32 | ND | >32 | ND | NS |
Micafungin | 0.082 | 0.016 | 0.113 | 0.048 | NS |
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Staerck, C.; Yaakoub, H.; Vandeputte, P.; Tabiasco, J.; Godon, C.; Gastebois, A.; Giraud, S.; Guillemette, T.; Calenda, A.; Delneste, Y.; et al. The Glycosylphosphatidylinositol-Anchored Superoxide Dismutase of Scedosporium apiospermum Protects the Conidia from Oxidative Stress. J. Fungi 2021, 7, 575. https://doi.org/10.3390/jof7070575
Staerck C, Yaakoub H, Vandeputte P, Tabiasco J, Godon C, Gastebois A, Giraud S, Guillemette T, Calenda A, Delneste Y, et al. The Glycosylphosphatidylinositol-Anchored Superoxide Dismutase of Scedosporium apiospermum Protects the Conidia from Oxidative Stress. Journal of Fungi. 2021; 7(7):575. https://doi.org/10.3390/jof7070575
Chicago/Turabian StyleStaerck, Cindy, Hajar Yaakoub, Patrick Vandeputte, Julie Tabiasco, Charlotte Godon, Amandine Gastebois, Sandrine Giraud, Thomas Guillemette, Alphonse Calenda, Yves Delneste, and et al. 2021. "The Glycosylphosphatidylinositol-Anchored Superoxide Dismutase of Scedosporium apiospermum Protects the Conidia from Oxidative Stress" Journal of Fungi 7, no. 7: 575. https://doi.org/10.3390/jof7070575