Cell ATP level of Saccharomyces cerevisiae sensitively responds to culture growth and drug-inflicted variations in membrane integrity and PDR pump activity

Krasowska, Anna; Łukaszewicz, Marcin; Bartosiewicz, Dominika; Sigler, Karel

doi:10.1016/j.bbrc.2010.03.133

Cited by 18 publications

(8 citation statements)

References 41 publications

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“…The reason for this difference in the rate of staining could be a lower PMP in C. albicans cells relative to S. cerevisiae cells. In the S. cerevisiae US 50–18C strain, with an overexpression of major pumps Pdr5p, Snq2p, and Yor1p, the cell ATP level varies depending on the growth phase and activity of PDR pumps ( Krasowska et al, 2010 ). This is not the case in C. albicans .…”

Section: Discussionmentioning

confidence: 99%

Detection of inhibitors of Candida albicans Cdr transporters using a diS-C3(3) fluorescence

2015

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Candida albicans is a major cause of opportunistic and life-threatening, systemic fungal infections. Hence new antifungal agents, as well as new methods to treat fungal infections, are still needed. The application of inhibitors of drug-efflux pumps may increase the susceptibility of C. albicans to drugs. We developed a new fluorescence method that allows the in vivo activity evaluation of compounds inhibiting of C. albicans transporters. We show that the potentiometric dye 3,3′-dipropylthiacarbocyanine iodide diS-C3(3) is pumped out by both Cdr1 and Cdr2 transporters. The fluorescence labeling with diS-C3(3) enables a real-time observation of the activity of C. albicans Cdr1 and Cdr2 transporters. We demonstrate that enniatin A and beauvericin show different specificities toward these transporters. Enniatin A inhibits diS-C3(3) efflux by Cdr1 while beauvericin inhibits both Cdr1p and Cdr2p.

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Section: Discussionmentioning

confidence: 99%

Detection of inhibitors of Candida albicans Cdr transporters using a diS-C3(3) fluorescence

2015

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“…Separating the cell growth phase and transporter expression phase by using phase-dependent promoters is an ideal strategy to alleviate the metabolic burden caused by transporter overexpression. Secondly, ABC transporters are ATP-based transporters, which hydrolyzing ATP to drive transport [20]. Therefore, sufficient ATP has to be supplied to the ABC transporter overexpressed strains.…”

Section: Introductionmentioning

confidence: 99%

Engineering endogenous ABC transporter with improving ATP supply and membrane flexibility enhances the secretion of β-carotene in Saccharomyces cerevisiae

Lin

Cheng

et al. 2020

Biotechnol Biofuels

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Background Product toxicity is one of the bottlenecks for microbial production of biofuels, and transporter-mediated biofuel secretion offers a promising strategy to solve this problem. As a robust microbial host for industrial-scale production of biofuels, Saccharomyces cerevisiae contains a powerful transport system to export a wide range of toxic compounds to sustain survival. The aim of this study is to improve the secretion and production of the hydrophobic product (β-carotene) by harnessing endogenous ABC transporters combined with physiological engineering in S. cerevisiae. Results Substrate inducibility is a prominent characteristic of most endogenous transporters. Through comparative proteomic analysis and transcriptional confirmation, we identified five potential ABC transporters (Pdr5p, Pdr10p, Snq2p, Yor1p, and Yol075cp) for β-carotene efflux. The accumulation of β-carotene also affects cell physiology in various aspects, including energy metabolism, mitochondrial translation, lipid metabolism, ergosterol biosynthetic process, and cell wall synthesis. Here, we adopted an inducible GAL promoter to overexpress candidate transporters and enhanced the secretion and intracellular production of β-carotene, in which Snq2p showed the best performance (a 4.04-fold and a 1.33-fold increase compared with its parental strain YBX-01, respectively). To further promote efflux capacity, two strategies of increasing ATP supply and improving membrane fluidity were following adopted. A 5.80-fold increase of β-carotene secretion and a 1.71-fold increase of the intracellular β-carotene production were consequently achieved in the engineered strain YBX-20 compared with the parental strain YBX-01. Conclusions Overall, our results showcase that engineering endogenous plasma membrane ABC transporters is a promising approach for hydrophobic product efflux in S. cerevisiae. We also highlight the importance of improving cell physiology to enhance the efficiency of ABC transporters, especially energy status and cell membrane properties.

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“…Such a protection requires an energy investment because such pumps consume ATP. Accordingly, Saccharomyces cerevisiae cells with inactivated pleiotropic drug resistance (PDR) ABC transporters show increased ATP concentrations and reach stationary phase cell densities that are higher than those of the wild-type cells 3 . Moreover, under normal conditions, the overexpression of a major pleiotropic drug resistance ABC transporter PDR5 decreases the fitness of the strain 4 .…”

Section: Introductionmentioning

confidence: 99%

Penetrating cations induce pleiotropic drug resistance in yeast

Galkina

Besedina

Zinovkin

et al. 2018

Sci Rep

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Substrates of pleiotropic drug resistance (PDR) transporters can induce the expression of corresponding transporter genes by binding to their transcription factors. Penetrating cations are substrates of PDR transporters and theoretically may also activate the expression of transporter genes. However, the accumulation of penetrating cations inside mitochondria may prevent the sensing of these molecules. Thus, whether penetrating cations induce PDR is unclear. Using Saccharomyces cerevisiae as a model, we studied the effects of penetrating cations on the activation of PDR. We found that the lipophilic cation dodecyltriphenylphosphonium (C12TPP) induced the expression of the plasma membrane PDR transporter genes PDR5, SNQ2 and YOR1. Moreover, a 1-hour incubation with C12TPP increased the concentration of Pdr5p and Snq2p and prevented the accumulation of the PDR transporter substrate Nile red. The transcription factor PDR1 was required to mediate these effects, while PDR3 was dispensable. The deletion of the YAP1 or RTG2 genes encoding components of the mitochondria-to-nucleus signalling pathway did not prevent the C12TPP-induced increase in Pdr5-GFP. Taken together, our data suggest (i) that the sequestration of lipophilic cations inside mitochondria does not significantly inhibit sensing by PDR activators and (ii) that the activation mechanisms do not require mitochondria as a signalling module.

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Cell ATP level of Saccharomyces cerevisiae sensitively responds to culture growth and drug-inflicted variations in membrane integrity and PDR pump activity

Cited by 18 publications

References 41 publications

Detection of inhibitors of Candida albicans Cdr transporters using a diS-C3(3) fluorescence

Detection of inhibitors of Candida albicans Cdr transporters using a diS-C3(3) fluorescence

Engineering endogenous ABC transporter with improving ATP supply and membrane flexibility enhances the secretion of β-carotene in Saccharomyces cerevisiae

Penetrating cations induce pleiotropic drug resistance in yeast

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