Plasmodium falciparum Mitochondrial Complex III, the Target of Atovaquone, Is Essential for Progression to the Transmissible Sexual Stages
Abstract
:1. Introduction
2. Results
2.1. PfRieske and PfC3AP2 Are Part of Plasmodium falciparum Complex III (PfCIII)
2.2. PfRieske Is Essential for Parasite Growth
2.3. Loss of PfRieske Results in Hypersensitivity to mETC Inhibitors and to Membrane Depolarization by Proguanil
2.4. PfRieske Is Required for the Pyrimidine Synthesis Pathway
2.5. Loss of PfRieske Causes Decrease in Cristae Density and Affects Gametocytes Maturation
3. Discussion
4. Materials and Methods
4.1. Plasmid Construction, Parasite Culture, Transfection, and Transgenic Line Confirmation
4.2. Inducible Knockdown and Parasite Growth Assay
4.3. Immunoprecipitation and Mass Spectrometry
4.4. Denaturing Polyacrylamide Gel Electrophoresis (SDS-PAGE), Blue Native (BN-PAGE), and Western Blotting
4.5. Immuno-Fluorescence Assay (IFA) and Microscopy
4.6. Metabolomic Profiling
4.7. Drug Sensitivity Assay
4.8. Transmission Electron Microscopy
4.9. Gametocyte Commitment and Culturing
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Expected | Detected (Epx1) | Detected (Exp 2) | Accession |
---|---|---|---|
Cytb | Yes | Yes | PF3D7_MITO2300 |
Cytc1 | Yes | Yes | PF3D7_1462700 |
Rieske | Yes | Yes | PF3D7_1439400 |
QCR6 | No | No | PF3D7_1426900 |
QCR7 | Yes | Yes | PF3D7_1012300 |
QCR8 | No | No | PF3D7_0306000 |
QCR9 | No | Yes | PF3D7_0622600 |
MPPalpha | Yes | Yes | PF3D7_0523100 |
MPPbeta | Yes | Yes | PF3D7_0933600 |
C3AP1 | No | Yes | PF3D7_0722700 |
C3AP2 | Yes | Yes | PF3D7_1326000 |
C3AP3 | Yes | Yes | PF3D7_0817800 |
Name | Sequence |
---|---|
2448 PfC3AP2-FWD | GCAGATCTCCTTCAGATAGTAGGAAATTTG |
2449 PfC3AP2-REV | GCCTGCAGC TGCGTGTTCAACTGA |
2535 PfRieske-FWD-B | GCGGATCCCCTGTATTTCCAAGAAAACC |
2451 PfRieske-REV | GCCTGCAGCTCCAATTTTTATCGTATTTTC |
2718 PfC3AP2-Int1 | CAAAAGAATTCAGGGTAAATTTATACCTTG |
2719 PfRieske-Int1 | GTATTCCTCCTGCATCTGAAGATC |
2797 HAglmS-REV-1 | TGATCTGCACACTCAGCCGGGA |
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Sheokand, P.K.; Pradhan, S.; Maclean, A.E.; Mühleip, A.; Sheiner, L. Plasmodium falciparum Mitochondrial Complex III, the Target of Atovaquone, Is Essential for Progression to the Transmissible Sexual Stages. Int. J. Mol. Sci. 2024, 25, 9239. https://doi.org/10.3390/ijms25179239
Sheokand PK, Pradhan S, Maclean AE, Mühleip A, Sheiner L. Plasmodium falciparum Mitochondrial Complex III, the Target of Atovaquone, Is Essential for Progression to the Transmissible Sexual Stages. International Journal of Molecular Sciences. 2024; 25(17):9239. https://doi.org/10.3390/ijms25179239
Chicago/Turabian StyleSheokand, Pradeep Kumar, Sabyasachi Pradhan, Andrew E. Maclean, Alexander Mühleip, and Lilach Sheiner. 2024. "Plasmodium falciparum Mitochondrial Complex III, the Target of Atovaquone, Is Essential for Progression to the Transmissible Sexual Stages" International Journal of Molecular Sciences 25, no. 17: 9239. https://doi.org/10.3390/ijms25179239