Abstract
To understand the role of phospholipids on Cdr1p (drug exporter)-mediated drug resistance in yeast, the phospholipids biosynthesis genes PSD1, PSD2, CHO2, and OPI3 were deleted in a strain of Saccharomyces cerevisiae already overexpressing Cdr1-GFP of Candida albicans as a heterologous system. The effect of phospholipids biosynthesis gene deletion was analyzed on Cdr1p-GFP-mediated drug resistance as well as its localization. The results indicate that phospholipids biosynthesis disruption makes the cell sensitive to several drugs including fluconazole (FLC), with Δpsd1/Cdr1-GFP being worst affected. Interestingly, unlike sterols and sphingolipids, the localization of Cdr1p was unaffected by phospholipid biosynthesis gene disruption. Concomitantly, phospholipids mutants also showed an increase in reactive oxygen species (ROS) generation, as verified by fluorescence probe 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) method. In addition, the sensitivity of phospholipid mutants with FLC was found to be synergistic to ROS generation, resulting in further reduction of growth. Thus, this study proposes phospholipid biosynthesis as a novel target for antifungal therapy.
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Acknowledgments
We thank Prof. R. D. Cannon (University of Otago, New Zealand) and K. Natarajan (JNU, New Delhi) for providing us the plasmids and strains. We would like to acknowledge Dr. Garima Dalal (JNU, New Delhi) for her diligent efforts for language editing of this manuscript. The authors also thank Mr. Ashok Kumar Sahu from AIRF-JNU, for his support in confocal imaging.
Funding
This work was supported by the individual grant to R. Pasrija from University Grant Commission (F.No. 42-647(2013) SR); Department of Science and Technology (SERB/F/4213/2013-14), and Department of Biotechnology IPLS funding (BT/PR4329/INF’22/1444/2011) to Maharshi Dayanand University.
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Kundu, D., Hameed, S., Fatima, Z. et al. Phospholipid biosynthesis disruption renders the yeast cells sensitive to antifungals. Folia Microbiol 65, 121–131 (2020). https://doi.org/10.1007/s12223-019-00713-3
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DOI: https://doi.org/10.1007/s12223-019-00713-3