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Updated: Apr 25, 2026
Genetic Mapping of Thermotolerance Differences Between Species of Saccharomyces Yeast via Genome-Wide Reciprocal Hemizygosity Analysis
Published on: August 12, 2019
Genetic analysis of resistance and sensitivity to 2-deoxyglucose in Saccharomyces cerevisiae
Rhonda R McCartney1, Dakshayini G Chandrashekarappa1, Bob B Zhang1
1Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15261.
Abstract:
Aerobic glycolysis is a metabolic pathway utilized by human cancer cells and also by yeast cells when they ferment glucose to ethanol. Both cancer cells and yeast cells are inhibited by the presence of low concentrations of 2-deoxyglucose (2DG). Genetic screens in yeast used resistance to 2-deoxyglucose to identify a small set of genes that function in regulating glucose metabolism. A recent high throughput screen for 2-deoxyglucose resistance identified a much larger set of seemingly unrelated genes. Here, we demonstrate that these newly identified genes do not in fact confer significant resistance to 2-deoxyglucose. Further, we show that the relative toxicity of 2-deoxyglucose is carbon source dependent, as is the resistance conferred by gene deletions. Snf1 kinase, the AMP-activated protein kinase of yeast, is required for 2-deoxyglucose resistance in cells growing on glucose. Mutations in the SNF1 gene that reduce kinase activity render cells hypersensitive to 2-deoxyglucose, while an activating mutation in SNF1 confers 2-deoxyglucose resistance. Snf1 kinase activated by 2-deoxyglucose does not phosphorylate the Mig1 protein, a known Snf1 substrate during glucose limitation. Thus, different stimuli elicit distinct responses from the Snf1 kinase.
Insights
2-deoxyglucose (2DG) inhibits both cancer and yeast cells. Newly identified genes do not confer significant 2DG resistance, and Snf1 kinase activity is crucial for 2DG resistance in yeast, showing distinct kinase responses to different stimuli.
Area of Science:
- Biochemistry
- Cell Biology
- Metabolic Regulation
Background:
- Aerobic glycolysis is a metabolic pathway used by cancer and yeast cells.
- 2-deoxyglucose (2DG) inhibits both cancer and yeast cells, making it a target for studying glucose metabolism.
- Previous genetic screens identified genes regulating glucose metabolism via 2DG resistance.
Purpose of the Study:
- To re-evaluate genes identified in a recent high-throughput screen for 2-deoxyglucose resistance.
- To investigate the role of Snf1 kinase in 2-deoxyglucose resistance.
- To understand the distinct responses of Snf1 kinase to different stimuli.
Main Methods:
- Genetic screens in yeast to identify 2-deoxyglucose resistance genes.
- Assessment of 2-deoxyglucose toxicity and resistance across different carbon sources.
- Analysis of Snf1 kinase activity and its effect on Mig1 phosphorylation.
Main Results:
- Newly identified genes from a high-throughput screen did not confer significant 2-deoxyglucose resistance.
- The toxicity of 2-deoxyglucose and resistance from gene deletions are carbon source-dependent.
- Snf1 kinase is essential for 2-deoxyglucose resistance on glucose; mutations affect sensitivity, and activation confers resistance.
- Activated Snf1 kinase does not phosphorylate Mig1 in response to 2-deoxyglucose.
Conclusions:
- The previously identified large set of genes for 2DG resistance may not be valid.
- Snf1 kinase plays a critical role in mediating 2-deoxyglucose resistance in yeast.
- Snf1 kinase exhibits stimulus-specific regulatory mechanisms, distinct from its response to glucose limitation.
