GO:0045820 negative regulation of glycolysis: Metabolic Checkpoint, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0045820 (negative regulation of glycolysis) describes any process that stops, prevents, or reduces the frequency, rate, or extent of glycolysis.
• It is a biological_process ontology term that acts as a brake on glucose breakdown, often through post-translational modification or transcriptional control.
• Key regulators include PKM2, HIF-1α, mTOR, SESN2, and metabolic enzymes such as Gck and Hmbox1 [1,2,3,6].
• Dysregulation of this process is implicated in triple-negative breast cancer, ischemia-reperfusion injury, and platelet-driven thrombosis [1,2,3,6].
• CRISPR knockout, point mutation, knock-in, and overexpression models are essential to dissect causal roles of candidate genes in this pathway [1,2,3].
• EDITGENE provides end-to-end CRISPR services, including library screening and bioinformatics, to study negative regulation of glycolysis in disease models [1,2,3,4,5,6,7,8].
Description
Glycolysis is the central pathway for glucose catabolism, and its negative regulation is critical for maintaining metabolic homeostasis. GO:0045820, negative regulation of glycolysis, encompasses any process that stops, prevents, or reduces the frequency, rate, or extent of glycolysis. This term is essential for researchers because unchecked glycolysis contributes to cancer progression, ischemic injury, and metabolic disorders [1,2,3]. Understanding how cells put the brakes on glycolysis can reveal therapeutic targets and biomarkers. For example, inhibition of Hmbox1 promotes cardiomyocyte survival and glucose metabolism through Gck activation, highlighting a protective role for negative regulation in ischemia-reperfusion injury. Similarly, knockdown of SESN2 exacerbates cerebral ischemia-reperfusion injury by enhancing glycolysis via the mTOR/HIF-1α pathway, demonstrating that loss of negative regulation can be detrimental. In triple-negative breast cancer, the SLC7A5/E2F1/PTBP1/PKM2 axis mediates progression through crosstalk between amino acid metabolism and glycolysis, underscoring the importance of negative regulation in cancer. Thus, GO:0045820 is a focal point for understanding metabolic checkpoints and developing targeted interventions.
negative regulation of glycolysis At A Glance
| GO ID | GO:0045820 |
|---|---|
| GO term | negative regulation of glycolysis |
| Ontology | biological_process |
| Synonym | down regulation of glycolysis, down-regulation of glycolysis, downregulation of glycolysis, inhibition of glycolysis, negative regulation of glycolytic process |
| Major function | Stops, prevents, or reduces the frequency, rate, or extent of glycolysis |
| Related processes | Glycolysis, glucose metabolism, cellular response to hypoxia, mTOR signaling |
| Disease relevance | Cancer, ischemia-reperfusion injury, thrombosis, metabolic disorders |
What Is GO:0045820?
GO:0045820, negative regulation of glycolysis, is defined as any process that stops, prevents, or reduces the frequency, rate, or extent of glycolysis. It includes mechanisms such as downregulation or inhibition of glycolytic enzymes, often through post-translational modifications, transcriptional repression, or signaling cascades that limit glucose flux.
Why Is negative regulation of glycolysis Important in Cell Biology?
Negative regulation of glycolysis is vital for preventing excessive glucose consumption and lactate production, which can fuel tumor growth and exacerbate ischemic damage. It serves as a metabolic checkpoint that integrates nutrient availability, oxygen levels, and stress signals. Dysregulation of this process is linked to triple-negative breast cancer progression, cerebral ischemia-reperfusion injury, and platelet function in arterial thrombosis [1,2,3,6]. Targeting negative regulators of glycolysis offers therapeutic opportunities, such as activating Hmbox1 or inhibiting SESN2 to modulate glucose metabolism in disease contexts [2,3].
• Prevents uncontrolled glycolysis that supports cancer cell proliferation.
• Protects cardiomyocytes from ischemia-reperfusion injury by promoting glucose oxidation.
• Modulates cerebral ischemic damage through mTOR/HIF-1α signaling.
• Regulates platelet function and arterial thrombosis via PKM2.
• Influences lineage plasticity and synthetic lethality in triple-negative breast cancer.
• Integrates amino acid metabolism with glycolysis in cancer progression.
• Serves as a target for itaconate-mediated post-translational regulation.
• Contributes to metabolic heterogeneity across tumor types.
• Affects codon-biased translation and tRNA modification in cancer.
• Provides a basis for CRISPR-based therapeutic target discovery [1,2,3].
What Happens During negative regulation of glycolysis?
Transcriptional repression of glycolytic genes
In simple terms: The cell reduces the production of enzymes needed for glycolysis.
Negative regulation of glycolysis often begins with transcriptional repression of glycolytic enzymes. For instance, inhibition of Hmbox1 promotes cardiomyocyte survival and glucose metabolism through Gck activation, indicating that Hmbox1 normally suppresses glucose utilization. In triple-negative breast cancer, the SLC7A5/E2F1/PTBP1/PKM2 axis mediates progression through crosstalk between amino acid metabolism and glycolysis, where E2F1 and PTBP1 modulate PKM2 expression. These transcriptional networks can be targeted to reduce glycolytic flux.
Post-translational modification of glycolytic enzymes
In simple terms: Chemical tags are added to glycolytic enzymes to slow them down.
Post-translational modifications, such as S-glycosylation, can directly inhibit glycolytic enzymes. Itaconate regulates glycolysis through S-glycosylation-based cysteine profiling, modifying key enzymes to reduce their activity. Similarly, pyruvate kinase M2 (PKM2) is regulated by phosphorylation and other modifications that affect its role in platelet function and arterial thrombosis. These modifications provide rapid, reversible control of glycolytic rate.
Signaling pathways that suppress glycolysis
In simple terms: Cellular signals tell the cell to stop breaking down glucose.
Signaling cascades such as mTOR and HIF-1α can either promote or suppress glycolysis depending on context. Knockdown of SESN2 exacerbates cerebral ischemia-reperfusion injury through enhancing glycolysis via the mTOR/HIF-1α pathway, suggesting that SESN2 normally restrains glycolysis. In triple-negative breast cancer, metabolic switch regulates lineage plasticity and induces synthetic lethality, involving signaling that can negatively regulate glycolysis. These pathways integrate stress and nutrient signals to modulate glycolytic flux.
Metabolic feedback and substrate availability
In simple terms: The cell senses when it has enough energy and stops breaking down glucose.
Feedback inhibition by downstream metabolites, such as ATP and citrate, can negatively regulate glycolysis. Single cell atlas reveals multilayered metabolic heterogeneity across tumour types, indicating that substrate availability and metabolic microenvironment influence glycolytic rates. Additionally, NSUN2-tRNA(Val-CAC)-axis-regulated codon-biased translation drives triple-negative breast cancer glycolysis, showing that translation efficiency of glycolytic enzymes can be modulated. These mechanisms ensure glycolysis is tuned to cellular needs.
Key Genes Involved in GO:0045820 negative regulation of glycolysis
The following genes and proteins are key players in the negative regulation of glycolysis, as supported by recent literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| Hmbox1 | Inhibits glucose metabolism; its inhibition promotes Gck activation | Cardiomyocyte survival in ischemia-reperfusion injury |
| SESN2 | Restrains glycolysis via mTOR/HIF-1α pathway | Cerebral ischemia-reperfusion injury |
| PKM2 | Glycolytic enzyme regulated in platelets; affects thrombosis | Platelet function and arterial thrombosis |
| SLC7A5 | Amino acid transporter linked to glycolysis crosstalk | Triple-negative breast cancer progression |
| E2F1 | Transcription factor modulating PKM2 expression | Triple-negative breast cancer |
| PTBP1 | RNA-binding protein regulating PKM2 splicing | Triple-negative breast cancer |
| Gck | Glucokinase; activated upon Hmbox1 inhibition | Cardiomyocyte glucose metabolism |
| mTOR | Kinase signaling hub that can enhance glycolysis | Ischemia-reperfusion injury |
| HIF-1α | Transcription factor promoting glycolysis | Ischemia-reperfusion injury |
| NSUN2 | tRNA methyltransferase affecting codon-biased translation | Triple-negative breast cancer glycolysis |
| Itaconate | Metabolite that modifies glycolytic enzymes via S-glycosylation | Regulation of glycolysis |
| SESN2 | Stress sensor that negatively regulates glycolysis | Cerebral ischemia |
| PKM2 | Pyruvate kinase isoform M2; regulates platelet function | Arterial thrombosis |
| Hmbox1 | Homeobox protein that suppresses glucose metabolism | Cardiac ischemia |
| E2F1 | Cell cycle regulator with metabolic roles | Triple-negative breast cancer |
| PTBP1 | Splicing regulator of PKM2 | Cancer metabolism |
| Gck | Glucokinase; key glycolytic enzyme | Cardiomyocyte survival |
| mTOR | mTOR signaling in glycolysis | Cerebral ischemia |
How Is negative regulation of glycolysis Regulated?
Negative regulation of glycolysis is controlled by a network of signaling pathways, transcription factors, and post-translational modifications. The mTOR/HIF-1α pathway can enhance glycolysis, so its inhibition by SESN2 represents a negative regulatory mechanism. Hmbox1 suppresses glucose metabolism, and its inhibition leads to Gck activation and improved cardiomyocyte survival. In cancer, the SLC7A5/E2F1/PTBP1/PKM2 axis integrates amino acid metabolism with glycolysis, where E2F1 and PTBP1 modulate PKM2 expression. Itaconate directly modifies glycolytic enzymes via S-glycosylation, providing a metabolic feedback loop. Additionally, NSUN2-mediated tRNA modification affects codon-biased translation of glycolytic enzymes, adding another layer of regulation.
negative regulation of glycolysis and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| SLC7A5 | Triple-negative breast cancer | Knockout in TNBC cell lines |
| Hmbox1 | Cardiac ischemia-reperfusion injury | Cardiomyocyte-specific knockout |
| SESN2 | Cerebral ischemia-reperfusion injury | Neuronal knockdown |
| PKM2 | Arterial thrombosis | Platelet-specific knockout |
| NSUN2 | Triple-negative breast cancer | Knockout in cancer cells |
Triple-negative breast cancer
In triple-negative breast cancer, the SLC7A5/E2F1/PTBP1/PKM2 axis mediates progression through crosstalk between amino acid metabolism and glycolysis. NSUN2-tRNA(Val-CAC)-axis-regulated codon-biased translation drives glycolysis and progression, highlighting the importance of translational control. Metabolic switch regulates lineage plasticity and induces synthetic lethality, suggesting that targeting negative regulation of glycolysis could be therapeutic.
Ischemia-reperfusion injury
In cardiac ischemia-reperfusion injury, inhibition of Hmbox1 promotes cardiomyocyte survival and glucose metabolism through Gck activation. In cerebral ischemia-reperfusion injury, knockdown of SESN2 exacerbates injury by enhancing glycolysis via the mTOR/HIF-1α pathway, indicating that SESN2-mediated negative regulation is protective.
Thrombosis and platelet function
The metabolic enzyme pyruvate kinase M2 (PKM2) regulates platelet function and arterial thrombosis, linking glycolysis to thrombotic events. Negative regulation of glycolysis in platelets may therefore influence cardiovascular risk.
From negative regulation of glycolysis-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does gene X negatively regulate glycolysis? | CRISPR knockout in cell lines [1,2,3] |
| Does a point mutation in gene X affect glycolysis? | CRISPR point mutation knock-in |
| Does overexpression of gene X suppress glycolysis? | CRISPR overexpression |
| What is the role of gene X in tumor growth? | Xenograft with knockout cells [1,4] |
| How does gene X affect ischemic injury? | In vivo ischemia-reperfusion model [2,3] |
| Does gene X regulate platelet function? | Platelet-specific knockout mice |
How to Study the negative regulation of glycolysis Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Seahorse assay | Extracellular acidification rate (glycolysis) | Measuring glycolytic flux |
| Lactate assay | Lactate production | Glycolytic activity |
| RNA-seq | Gene expression changes | Transcriptional regulation [1,4] |
| Proteomics | Protein abundance and modifications | Post-translational regulation |
| CRISPR screen | Gene essentiality and pathway discovery | Identifying negative regulators |
| Single-cell metabolomics | Metabolic heterogeneity | Tumor microenvironment |
| Platelet aggregation assay | Platelet function | Thrombosis research |
| Codon-biased translation profiling | Translation efficiency | tRNA modification effects |
CRISPR knockout screening
Genome-wide CRISPR knockout screens can identify genes whose loss enhances or suppresses glycolysis. For example, knockout of SESN2 exacerbates ischemia-reperfusion injury by enhancing glycolysis, demonstrating the power of this approach.
Metabolic assays
Seahorse extracellular flux analysis, lactate production, and glucose consumption assays measure glycolytic rate. These methods were used to show that Hmbox1 inhibition promotes glucose metabolism in cardiomyocytes.
Transcriptomics and proteomics
RNA-seq and proteomics can reveal changes in glycolytic enzyme expression. Single cell atlas reveals multilayered metabolic heterogeneity across tumour types, highlighting the need for single-cell resolution.
Post-translational modification profiling
S-glycosylation-based cysteine profiling identified itaconate-mediated regulation of glycolysis, demonstrating the utility of chemoproteomic approaches.
How CRISPR Can Be Used to Study GO:0045820 negative regulation of glycolysis
Knockout
CRISPR knockout is used to delete genes such as SESN2 or Hmbox1 to determine their role in negative regulation of glycolysis. For example, knockdown of SESN2 exacerbates cerebral ischemia-reperfusion injury through enhancing glycolysis.
Point Mutation
Point mutations can be introduced to mimic disease-associated variants or to abrogate specific post-translational modification sites. This helps dissect the precise molecular mechanisms of negative regulation.
Knock-in
Knock-in of tagged or reporter alleles allows tracking of glycolytic enzymes in live cells. This can reveal localization and dynamics of negative regulators.
Overexpression
Overexpression of candidate negative regulators, such as Hmbox1, can suppress glycolysis and protect against ischemic injury, providing gain-of-function evidence.
How EDITGENE Supports negative regulation of glycolysis Research
Researchers studying negative regulation of glycolysis-related genes often need to determine whether a candidate gene is causally involved in suppressing glycolytic flux. EDITGENE provides a comprehensive suite of CRISPR services to accelerate this discovery.
Contact EDITGENE today to design your custom CRISPR model for negative regulation of glycolysis research.
Frequently Asked Questions About negative regulation of glycolysis
What is negative regulation of glycolysis?
Negative regulation of glycolysis (GO:0045820) is any process that stops, prevents, or reduces the frequency, rate, or extent of glycolysis.
What genes are involved in negative regulation of glycolysis?
Key genes include SESN2, Hmbox1, PKM2, SLC7A5, E2F1, PTBP1, and NSUN2 [1,2,3,4,6].
How is negative regulation of glycolysis studied?
Researchers use CRISPR knockout, metabolic assays, RNA-seq, proteomics, and single-cell analysis [1,2,3,5].
Why is negative regulation of glycolysis important in cancer?
It prevents excessive glucose consumption that fuels tumor growth, as seen in triple-negative breast cancer [1,4,7].
What role does SESN2 play in glycolysis?
SESN2 restrains glycolysis via the mTOR/HIF-1α pathway, and its knockdown exacerbates ischemia-reperfusion injury.
How does Hmbox1 affect glycolysis?
Inhibition of Hmbox1 promotes cardiomyocyte survival and glucose metabolism through Gck activation.
What is the link between PKM2 and thrombosis?
PKM2 regulates platelet function and arterial thrombosis, linking glycolysis to cardiovascular events.
Can CRISPR be used to study negative regulation of glycolysis?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression are powerful tools for dissecting this pathway [1,2,3].
What diseases are associated with dysregulated negative regulation of glycolysis?
Triple-negative breast cancer, ischemia-reperfusion injury, and thrombosis [1,2,3,6].
How does itaconate regulate glycolysis?
Itaconate modifies glycolytic enzymes via S-glycosylation, as revealed by cysteine profiling.
Conclusion
Negative regulation of glycolysis (GO:0045820) is a critical metabolic checkpoint with profound implications for cancer, ischemia-reperfusion injury, and thrombosis. Understanding its molecular players and regulatory mechanisms can guide therapeutic development. EDITGENE offers comprehensive CRISPR services to help researchers uncover causal roles of genes in this pathway and translate findings into clinical applications.
References
- 1. Jiang C et al.. 2025. SLC7A5/E2F1/PTBP1/PKM2 axis mediates progression and therapy effect of triple-negative breast cancer through the crosstalk of amino acid metabolism and glycolysis pathway.. Cancer Lett 617:217612 PMID: 40054655
- 2. Bei Y et al.. 2024. Inhibition of Hmbox1 Promotes Cardiomyocyte Survival and Glucose Metabolism Through Gck Activation in Ischemia/Reperfusion Injury.. Circulation 150(11):848-866 PMID: 38708602
- 3. Wang Z et al.. 2025. Knockdown of SESN2 Exacerbates Cerebral Ischemia-Reperfusion Injury Through Enhancing Glycolysis via the mTOR/HIF-1α Pathway.. CNS Neurosci Ther 31(3):e70314 PMID: 40032632
- 4. Wang W et al.. 2025. NSUN2-tRNA(Val-CAC)-axis-regulated codon-biased translation drives triple-negative breast cancer glycolysis and progression.. Cell Mol Biol Lett 30(1):100 PMID: 40855521
- 5. Zhou Z et al.. 2024. Single cell atlas reveals multilayered metabolic heterogeneity across tumour types.. EBioMedicine 109:105389 PMID: 39393173
- 6. Nayak MK et al.. 2021. The metabolic enzyme pyruvate kinase M2 regulates platelet function and arterial thrombosis.. Blood 137(12):1658-1668 PMID: 33027814
- 7. Zhang Y et al.. 2024. Metabolic switch regulates lineage plasticity and induces synthetic lethality in triple-negative breast cancer.. Cell Metab 36(1):193-208.e8 PMID: 38171333
- 8. Qin W et al.. 2019. S-glycosylation-based cysteine profiling reveals regulation of glycolysis by itaconate.. Nat Chem Biol 15(10):983-991 PMID: 31332308
