A catabolic block does not sufficiently explain how 2-deoxy-D-glucose inhibits cell growth - PubMed (original) (raw)

A catabolic block does not sufficiently explain how 2-deoxy-D-glucose inhibits cell growth

Markus Ralser et al. Proc Natl Acad Sci U S A. 2008.

Abstract

The glucose analogue 2-deoxy-D-glucose (2-DG) restrains growth of normal and malignant cells, prolongs the lifespan of C. elegans, and is widely used as a glycolytic inhibitor to study metabolic activity with regard to cancer, neurodegeneration, calorie restriction, and aging. Here, we report that separating glycolysis and the pentose phosphate pathway highly increases cellular tolerance to 2-DG. This finding indicates that 2-DG does not block cell growth solely by preventing glucose catabolism. In addition, 2-DG provoked similar concentration changes of sugar-phosphate intermediates in wild-type and 2-DG-resistant yeast strains and in human primary fibroblasts. Finally, a genome-wide analysis revealed 19 2-DG-resistant yeast knockouts of genes implicated in carbohydrate metabolism and mitochondrial homeostasis, as well as ribosome biogenesis, mRNA decay, transcriptional regulation, and cell cycle. Thus, processes beyond the metabolic block are essential for the biological properties of 2-DG.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Fig. 1.

Yeast cells interrupted for the interplay of glycolysis and the PPP are 2-DG resistant. (A) Simplified overview of glycolysis, the PPP, and the enzymatic targets of 2-DG. (B) Wild-type and Δ_ZWF1_ yeast were spotted as 5-fold serial dilution on SC media with or without 2-DG and incubated at 30 °C. (C) Similar to (B), but with yeast cells expressing K. lactis GDP1. (D) Wild-type and Δ_ZWF1_ yeast transformed with a K. lactis GDP1 expression plasmid were spotted on SC-URA media with or without 2-DG

Fig. 2.

Metabolic profiling of eukaryotic cells treated with 2-DG. (A) Growth of yeast batch cultures supplemented with 2-DG at different concentrations. (B) Quantitative changes of metabolites in wild-type, Δ_ZWF1_, and _GDP1_-expressing yeast treated with 2-DG. Shown are the relative increases or decreases in percentage compared with the respective untreated strain. ◇, e4p was below detection limit in the 2-DG-untreated cells, but strongly increased (to 6.5 ± 0.55 mM/(OD600 × ml) [WT yeast], 9.9 ± 0.76 mM/(OD600 × ml) [Δ_ZWF1_], and 6.5 ± 0.67 mM/(OD600 × ml) [_GDP1_] in the 2-DG-treated sample). (C) Human primary fibroblasts were incubated with 4 mM 2-DG and processed for sugar-phosphate analyses. ◇, e4p increased to 9.70 nmol/mgtotal protein. (B and C) Error bars indicate the normalized standard deviation of the 2-DG-treated cells.

Fig. 3.

Identification of 2-DG-resistant yeast knockouts. 2-DG-resistant yeast strains identified in the genome-wide screen were spotted, alternating with the wild-type strain, on SC media with or without 2-DG, as indicated. Growth was monitored after incubation for 3 days at 30°C.

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A catabolic block does not sufficiently explain how 2-deoxy-D-glucose inhibits cell growth - PubMed (original) (raw)