Screening the yeast genome for energetic metabolism pathways involved in a phenotypic response to the anti-cancer agent 3-bromopyruvate.

Paweł Lis,Stanisław Ułaszewski,Magdalena Cal-Bąkowska,Andre Goffeau,Paweł Jurkiewicz,Peter L Pedersen,Young H Ko

doi:10.18632/oncotarget.7174

Paweł Lis, Stanisław Ułaszewski + Show 5 more

Open Access

https://doi.org/10.18632/oncotarget.7174

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Abstract

In this study the detailed characteristic of the anti-cancer agent 3-bromopyruvate (3-BP) activity in the yeast Saccharomyces cerevisiae model is described, with the emphasis on its influence on energetic metabolism of the cell. It shows that 3-BP toxicity in yeast is strain-dependent and influenced by the glucose-repression system. Its toxic effect is mainly due to the rapid depletion of intracellular ATP. Moreover, lack of the Whi2p phosphatase results in strongly increased sensitivity of yeast cells to 3-BP, possibly due to the non-functional system of mitophagy of damaged mitochondria through the Ras-cAMP-PKA pathway. Single deletions of genes encoding glycolytic enzymes, the TCA cycle enzymes and mitochondrial carriers result in multiple effects after 3-BP treatment. However, it can be concluded that activity of the pentose phosphate pathway is necessary to prevent the toxicity of 3-BP, probably due to the fact that large amounts of NADPH are produced by this pathway, ensuring the reducing force needed for glutathione reduction, crucial to cope with the oxidative stress. Moreover, single deletions of genes encoding the TCA cycle enzymes and mitochondrial carriers generally cause sensitivity to 3-BP, while totally inactive mitochondrial respiration in the rho0 mutant resulted in increased resistance to 3-BP.

Highlights

Research using yeast Saccharomyces cerevisiae has provided much information concerning the mechanisms of cellular metabolic processes, cell cycle regulation, recombination, replication and repair of DNA as well as the cell death mechanisms, protein folding and biogenesis of organelles [1]
It can be concluded that activity of the pentose phosphate pathway is necessary to prevent the toxicity of 3-BP, probably due to the fact that large amounts of NADPH are produced by this pathway, ensuring the reducing force needed for glutathione reduction, crucial to cope with the oxidative stress
The metabolism of pyruvate in yeast and mammalian cells is not identical, still the general mechanism is the same – increased flux of this pathway diminishes the availability of substrates for aerobic metabolism, as it is in the case of hyperactive lactate dehydrogenase (LDH) in cancer cells

Summary

INTRODUCTION

Research using yeast Saccharomyces cerevisiae has provided much information concerning the mechanisms of cellular metabolic processes, cell cycle regulation, recombination, replication and repair of DNA as well as the cell death mechanisms, protein folding and biogenesis of organelles [1]. The metabolism of pyruvate in yeast and mammalian cells is not identical, still the general mechanism is the same – increased flux of this pathway diminishes the availability of substrates for aerobic metabolism, as it is in the case of hyperactive LDH in cancer cells All these suggest that during intense fermentation, Pdc activation is strictly connected to the repression of mitochondrial respiration [19]. Research in breast cancer cells shown that butyrate, which induces the expression of monocarboxylate transporter MCT4, causes increased sensitivity to 3-BP [28] It was shown using the KBM7 human myeloid leukemia cell line, that MCT1 mediates the uptake of 3-BP and that deletion of the MCT1 results in resistance to 3-BP [29]. This is the first report showing multidirectional influence of 3-BP on the yeast Saccharomyces cerevisiae, www.impactjournals.com/oncotarget a very useful model system for genomic screen of deletion mutants defective in the energetic metabolism

RESULTS

DISCUSSION

MATERIALS AND METHODS