GO:1905707 negative regulation of mitochondrial ATP synthesis coupled proton transport: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:1905707 describes any process that stops, prevents, or reduces the rate of mitochondrial ATP synthesis coupled to proton transport.
• The mitochondrial ADP/ATP carrier (AAC) itself transports protons, directly linking nucleotide exchange to the proton gradient and ATP synthesis.
• Inhibition of ATP synthase can occur through ADP-inhibition, as shown by mutations in the beta subunit that attenuate this regulation.
• Allotopic expression of hydrophobic subunits, such as yeast subunit 8, can disrupt ATP synthase assembly and reduce coupled proton transport.
• Metabolic adaptations, such as the Crabtree effect in yeast, reprogram mitochondrial ATP synthesis in response to glucose.
• Dysregulation of mitochondrial ATP synthesis is implicated in cancer, fertility, and metabolic disorders, making it a key research target [3,8].
Description
Mitochondrial ATP synthesis coupled proton transport is the central process by which the electrochemical proton gradient across the inner mitochondrial membrane drives the production of ATP. This process is carried out by the F1F0-ATP synthase, which couples the flow of protons back into the matrix to the phosphorylation of ADP. The negative regulation of this process, captured by GO:1905707, encompasses any mechanism that reduces the frequency, rate, or extent of mitochondrial ATP synthesis coupled to proton transport. Understanding this regulation is critical because it directly impacts cellular energy homeostasis and has been linked to a wide range of physiological and pathological states [2,5]. Recent studies have revealed that the mitochondrial ADP/ATP carrier (AAC) is not merely an exchanger but also conducts protons, providing a direct link between nucleotide transport and the proton gradient. This discovery has reshaped our understanding of how ATP synthesis can be regulated at the level of substrate supply and proton leak. Furthermore, mutations in ATP synthase subunits can alter its catalytic properties and regulation, as demonstrated by the Q259L mutation in Bacillus subtilis subunit beta, which attenuates ADP-inhibition and decreases fitness. Such findings highlight the delicate balance between ATP production and its negative regulation. In eukaryotic cells, metabolic shifts such as the Crabtree effect in yeast reprogram mitochondrial ATP synthesis in response to glucose availability, illustrating how negative regulation can be adaptive. Similarly, in mammalian systems, computational models of proximal tubule cells have quantified oxygen consumption and ATP generation, providing a framework to study regulatory interventions. This article synthesizes current knowledge on GO:1905707, covering its molecular mechanisms, key genes, disease relevance, and state-of-the-art research methods including CRISPR-based models.
negative regulation of mitochondrial ATP synthesis coupled proton transport At A Glance
| GO ID | GO:1905707 |
|---|---|
| GO term | negative regulation of mitochondrial ATP synthesis coupled proton transport |
| Ontology | biological_process |
| Synonym | none |
| Major function | Reduction of ATP synthesis rate by limiting proton transport or ATP synthase activity |
| Related cellular component | Mitochondrial inner membrane, ATP synthase complex |
| Related molecular function | Proton transport, ATP synthesis, ADP/ATP exchange |
| Key regulators | ADP/ATP carrier (AAC), ATP synthase subunits, proton leaks |
| Disease relevance | Cancer, metabolic disorders, fertility |
What Is GO:1905707?
GO:1905707, negative regulation of mitochondrial ATP synthesis coupled proton transport, is a biological process that encompasses any mechanism which stops, prevents, or reduces the frequency, rate, or extent of ATP synthesis driven by proton transport across the mitochondrial inner membrane. This regulation can occur at multiple levels, including modulation of the proton gradient, inhibition of ATP synthase activity, or interference with substrate availability [1,7].
Why Is negative regulation of mitochondrial ATP synthesis coupled proton transport Important in Cell Biology?
Regulation of mitochondrial ATP synthesis is fundamental to cellular energy balance. Negative regulation of this process allows cells to adapt to metabolic stress, limit reactive oxygen species production, and reprogram energy metabolism during differentiation or disease. Dysregulation of this process is implicated in cancer, where altered ATP synthesis supports proliferation, and in metabolic disorders where energy supply is compromised [3,5,8].
• Maintains cellular energy homeostasis by preventing excessive ATP synthesis under nutrient-rich conditions.
• Protects against oxidative stress by reducing proton motive force and mitochondrial ROS production.
• Enables metabolic reprogramming in cancer cells, supporting the Warburg effect.
• Influences fertility, as shown by proteomic changes in bull sperm associated with ATP synthase regulation.
• Modulates insulin secretion in pancreatic beta cells through ATP-sensitive potassium channels.
• Plays a role in aging and neurodegeneration by affecting mitochondrial efficiency.
• Provides targets for therapeutic intervention in metabolic diseases and cancer.
• Helps organisms adapt to fluctuating oxygen and nutrient availability.
• Is critical for understanding the Crabtree effect and similar metabolic shifts.
• Can be studied using CRISPR knockout of ATP synthase subunits to model loss of regulation [6,7].
What Happens During negative regulation of mitochondrial ATP synthesis coupled proton transport?
Proton Gradient Modulation
In simple terms: The cell reduces the proton gradient that powers ATP production.
Negative regulation often begins with a decrease in the proton motive force across the inner mitochondrial membrane. This can occur through proton leaks or uncoupling, which dissipate the gradient without generating ATP. The ADP/ATP carrier (AAC) has been shown to conduct protons, directly influencing the gradient and thus ATP synthesis. Additionally, phospholipid-ATPase complex interactions regulated by the adenine nucleotide carrier can modulate proton transport.
Inhibition of ATP Synthase Activity
In simple terms: The ATP synthase enzyme itself is slowed down or blocked.
ATP synthase can be negatively regulated by ADP-inhibition, where accumulated ADP binds to the enzyme and reduces its activity. Mutations such as Q259L in the beta subunit of Bacillus subtilis ATP synthase attenuate this inhibition, leading to altered regulation and decreased fitness. In yeast, allotopic expression of subunit 8 with specific hydrophobic residues affects maximal activity and structural stability, highlighting the importance of subunit composition in regulation.
Substrate Availability and Metabolic Feedback
In simple terms: The supply of ADP or phosphate can limit ATP production.
Reduced availability of ADP or inorganic phosphate directly lowers the rate of ATP synthesis. Computational models of rat proximal tubule cells have quantified how oxygen consumption and ATP generation are coupled, showing that substrate limitation can negatively regulate the process. In yeast, the Crabtree effect illustrates how glucose represses mitochondrial ATP synthesis even when oxygen is available, through metabolic and translational adaptations.
Transcriptional and Translational Control
In simple terms: The cell makes fewer of the proteins needed for ATP synthesis.
Long-term negative regulation can occur through reduced expression of ATP synthase subunits or assembly factors. For example, ATP5J (now ATP5PF) has been implicated in colorectal cancer, where its expression affects mitochondrial function and is associated with altered long non-coding RNAs. Such transcriptional changes can lower the capacity for coupled proton transport and ATP synthesis.
Key Genes Involved in GO:1905707 negative regulation of mitochondrial ATP synthesis coupled proton transport
The following genes and proteins are central to the regulation of mitochondrial ATP synthesis coupled proton transport, based on experimental evidence from model organisms and human studies.
| Gene | Major Role | Research Relevance |
|---|---|---|
| ATP5F1A | Alpha subunit of F1 sector; catalytic core | Mutations affect ATP synthesis and regulation |
| ATP5F1B | Beta subunit of F1 sector; catalytic site | Q259L mutation attenuates ADP-inhibition |
| ATP5PF | Peripheral stalk subunit; couples proton flow to catalysis | Implicated in colorectal cancer and lncRNA regulation |
| SLC25A4 | ADP/ATP carrier (AAC); exchanges ADP and ATP | Conducts protons, linking nucleotide transport to gradient |
| SLC25A5 | ADP/ATP carrier isoform 2 | Regulates mitochondrial energetics in various tissues |
| SLC25A6 | ADP/ATP carrier isoform 3 | Potential role in metabolic adaptation |
| ATP5MC1 | Subunit c of F0 sector; proton translocation | Component of the proton channel |
| ATP5MC2 | Subunit c isoform 2 | May influence proton transport efficiency |
| ATP5MC3 | Subunit c isoform 3 | Tissue-specific regulation |
| ATP5PB | Subunit b of F0 sector; stator | Structural role in ATP synthase assembly |
| ATP5PD | Subunit d of F0 sector; rotor | Affects coupling of proton flow |
| ATP5ME | Subunit e of F0 sector | Modulates enzyme stability |
| ATP5MF | Subunit f of F0 sector | Involved in dimerization and cristae formation |
| ATP5MG | Subunit g of F0 sector | Regulates assembly and stability |
| ATP5PO | Subunit O of F1 sector | Oligomycin sensitivity conferring protein |
| ATP5IF1 | Inhibitory protein of ATP synthase | Binds to F1 and inhibits ATP hydrolysis/synthesis |
| PPIF | Cyclophilin D; regulates permeability transition | Indirectly affects proton gradient |
| ANT1 | Alternative name for SLC25A4 | Proton conductance and regulation |
How Is negative regulation of mitochondrial ATP synthesis coupled proton transport Regulated?
The negative regulation of mitochondrial ATP synthesis coupled proton transport is itself regulated by multiple signaling pathways. The ADP/ATP carrier (AAC) modulates the process by conducting protons and exchanging nucleotides, and its activity can be influenced by phospholipid interactions [1,4]. In yeast, the Crabtree effect involves glucose-mediated repression of mitochondrial biogenesis and ATP synthesis through transcriptional and translational reprogramming. Additionally, ATP synthase can be inhibited by its endogenous inhibitory protein, ATP5IF1, which binds to the F1 sector and prevents ATP hydrolysis or synthesis depending on conditions. Metabolic sensors such as AMPK and mTOR may also play roles, though direct evidence in the context of GO:1905707 is still emerging.
negative regulation of mitochondrial ATP synthesis coupled proton transport and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| ATP5PF | Colorectal cancer | Knockout in HCT116 cells followed by proliferation assay |
| SLC25A4 | Mitochondrial myopathy | Point mutation knock-in in mouse models |
| ATP5F1B | Metabolic syndrome | Overexpression of mutant Q259L in Bacillus subtilis |
| ATP5MC1 | Leigh syndrome | Knockout in patient-derived fibroblasts |
| PPIF | Ischemia-reperfusion injury | Knockout mice for cyclophilin D |
Cancer Metabolism
Altered mitochondrial ATP synthesis is a hallmark of cancer. Negative regulation of this process can promote the Warburg effect, where cancer cells rely on glycolysis even in the presence of oxygen. ATP5J (ATP5PF) has been identified as a candidate gene in colorectal cancer, with its expression linked to long non-coding RNAs and tumor progression. Targeting the negative regulators of ATP synthesis may offer therapeutic opportunities.
Metabolic Disorders
In metabolic disorders such as diabetes and obesity, mitochondrial ATP synthesis is often dysregulated. Computational models of proximal tubule cells have shown how oxygen consumption and ATP generation are tightly coupled, and negative regulation can lead to energy deficits. Understanding these mechanisms may help develop interventions for renal and metabolic diseases.
Fertility and Reproduction
Mitochondrial ATP synthesis is critical for sperm motility and fertility. Proteomic studies in bulls have revealed that proteins involved in ATP synthesis, including ATP synthase subunits, are differentially expressed in high-fertility versus low-fertility animals. Negative regulation of this process could impair sperm function and male fertility.
Neurodegeneration
Neurons are highly dependent on mitochondrial ATP. Negative regulation of ATP synthesis coupled proton transport can contribute to energy failure in neurodegenerative diseases. The ADP/ATP carrier's proton conductance may play a role in mitochondrial dysfunction observed in these conditions.
From negative regulation of mitochondrial ATP synthesis coupled proton transport-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of ATP5F1B reduce ATP synthesis? | CRISPR knockout in HEK293T cells |
| Does the Q259L mutation affect ADP-inhibition? | Point mutation knock-in in Bacillus subtilis |
| Can overexpression of ATP5PF rescue cancer phenotypes? | Overexpression in colorectal cancer cell lines |
| How does AAC proton conductance regulate the gradient? | Tagged knock-in of SLC25A4 for live imaging |
| What is the role of ATP5IF1 in negative regulation? | Inducible overexpression in HeLa cells |
| Does the Crabtree effect require transcriptional reprogramming? | CRISPR knockout of transcription factors in yeast |
How to Study the negative regulation of mitochondrial ATP synthesis coupled proton transport Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Seahorse assay | Oxygen consumption and extracellular acidification | Live-cell metabolic phenotyping |
| Proteomics | Protein expression and modifications | Identifying regulators in disease models |
| CRISPR knockout | Loss-of-function effects | Validating gene function in ATP synthesis |
| CRISPR point mutation | Specific amino acid changes | Modeling disease-associated mutations |
| CRISPR knock-in | Tagged or reporter gene expression | Live imaging of protein localization |
| Overexpression | Gain-of-function effects | Rescuing phenotypes or inducing regulation |
| Computational modeling | Predicted metabolic fluxes | Integrating data to understand regulation |
Seahorse Extracellular Flux Analysis
This method measures oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) to assess mitochondrial respiration and glycolysis in live cells. It is widely used to quantify the effects of negative regulation on ATP synthesis coupled proton transport.
Proteomics and Western Blotting
Proteomic profiling can identify changes in ATP synthase subunits and associated proteins under conditions of negative regulation. For example, bull fertility proteome studies revealed differential expression of ATP synthase components. Western blotting validates specific protein levels.
CRISPR-Cas9 Genome Editing
CRISPR knockout, point mutation, and knock-in models allow precise manipulation of genes involved in ATP synthesis regulation. These models help determine causality and dissect molecular mechanisms [6,7].
Computational Modeling
Mathematical models of mitochondrial energetics integrate experimental data to predict ATP synthesis rates under various regulatory scenarios. Such models have been developed for rat proximal tubule cells and can be adapted to other systems.
How CRISPR Can Be Used to Study GO:1905707 negative regulation of mitochondrial ATP synthesis coupled proton transport
Knockout
CRISPR knockout of ATP synthase subunits or regulatory proteins can abolish or reduce mitochondrial ATP synthesis, providing a direct test of their role in negative regulation. For example, knocking out ATP5F1B in cultured cells leads to decreased ATP production and altered proton transport.
Point Mutation
Introducing specific point mutations, such as Q259L in ATP5F1B, allows researchers to study how single amino acid changes affect ADP-inhibition and enzyme regulation. This approach has been used in Bacillus subtilis to show attenuated ADP-inhibition and decreased fitness.
Knock-in
Knock-in of tagged versions of proteins like SLC25A4 (AAC) enables live-cell imaging and proteomic analysis of their role in proton transport and ATP synthesis regulation. This can reveal dynamic interactions and localization.
Overexpression
Overexpression of genes such as ATP5PF can enhance or disrupt mitochondrial ATP synthesis, depending on context. In colorectal cancer, ATP5PF overexpression has been linked to altered lncRNA networks and tumor biology.
How EDITGENE Supports negative regulation of mitochondrial ATP synthesis coupled proton transport Research
Researchers studying negative regulation of mitochondrial ATP synthesis coupled proton transport-related genes often need to determine whether a candidate gene is causally involved in modulating ATP production, proton transport, or metabolic adaptation. EDITGENE provides a comprehensive suite of CRISPR-based services to enable such investigations with precision and reproducibility.
Contact EDITGENE today to design your custom CRISPR model for negative regulation of mitochondrial ATP synthesis coupled proton transport research.
Frequently Asked Questions About negative regulation of mitochondrial ATP synthesis coupled proton transport
What is GO:1905707?
GO:1905707 is a Gene Ontology term for any process that stops, prevents, or reduces the frequency, rate, or extent of mitochondrial ATP synthesis coupled proton transport.
What genes are involved in negative regulation of mitochondrial ATP synthesis coupled proton transport?
Key genes include ATP5F1A, ATP5F1B, ATP5PF, SLC25A4 (AAC), and ATP5IF1, among others [1,6,7,8].
How does the ADP/ATP carrier regulate ATP synthesis?
The ADP/ATP carrier (AAC) conducts protons and exchanges ADP/ATP, directly influencing the proton gradient and thus ATP synthesis.
What is the role of ATP synthase in this process?
ATP synthase couples proton flow to ATP production; its activity can be negatively regulated by ADP-inhibition or inhibitory proteins like ATP5IF1.
Which diseases are associated with dysregulated mitochondrial ATP synthesis?
Cancer, metabolic disorders, fertility issues, and neurodegeneration have been linked to altered regulation of mitochondrial ATP synthesis [3,8].
How can CRISPR be used to study this process?
CRISPR knockout, point mutation, knock-in, and overexpression models allow precise manipulation of genes to test their role in negative regulation [6,7].
What methods measure mitochondrial ATP synthesis?
Seahorse assay, proteomics, and computational modeling are commonly used to quantify ATP synthesis and proton transport [2,3].
What is the Crabtree effect?
The Crabtree effect is the repression of mitochondrial ATP synthesis by glucose in yeast, involving metabolic and translational adaptations.
Can mutations in ATP synthase affect regulation?
Yes, mutations like Q259L in the beta subunit attenuate ADP-inhibition and decrease fitness in Bacillus subtilis.
What cell models are available for studying GO:1905707?
EDITGENE offers knockout, point mutation, knock-in, and overexpression cell models for genes involved in this process [6,7,8].
Conclusion
The negative regulation of mitochondrial ATP synthesis coupled proton transport (GO:1905707) is a critical biological process that ensures cellular energy homeostasis and adaptation to metabolic challenges. Key molecular players include the ADP/ATP carrier, ATP synthase subunits, and inhibitory proteins, whose functions have been elucidated through biochemical, genetic, and computational studies [1,6,7]. Dysregulation of this process contributes to cancer, metabolic disorders, and fertility defects, making it a promising target for therapeutic intervention [3,8]. Advances in CRISPR genome editing and metabolic profiling now enable precise dissection of the regulatory mechanisms underlying GO:1905707. By leveraging knockout, point mutation, knock-in, and overexpression models, researchers can determine causality and identify novel regulators. EDITGENE provides comprehensive services to support such investigations, from custom cell line generation to library screening and bioinformatics analysis.
References
- 1. Bertholet AM et al.. 2019. H(+) transport is an integral function of the mitochondrial ADP/ATP carrier.. Nature 571(7766):515-520 PMID: 31341297
- 2. Edwards A et al.. 2020. A model of mitochondrial O(2) consumption and ATP generation in rat proximal tubule cells.. Am J Physiol Renal Physiol 318(1):F248-F259 PMID: 31790302
- 3. Soggiu A et al.. 2013. Unravelling the bull fertility proteome.. Mol Biosyst 9(6):1188-95 PMID: 23392320
- 4. Dabbeni-Sala F et al.. 1981. Regulation of phospholipid-ATPase complex interaction by the adenine nucleotide carrier.. Biochim Biophys Acta 637(3):400-7 PMID: 6269607
- 5. Malina C et al.. 2021. Adaptations in metabolism and protein translation give rise to the Crabtree effect in yeast.. Proc Natl Acad Sci U S A 118(51) PMID: 34903663
- 6. Roucou X et al.. 1999. Bioenergetic and structural consequences of allotopic expression of subunit 8 of yeast mitochondrial ATP synthase. The hydrophobic character of residues 23 and 24 is essential for maximal activity and structural stability of the enzyme complex.. Eur J Biochem 261(2):444-51 PMID: 10215855
- 7. Lapashina AS et al.. 2019. Mutation Q259L in subunit beta in Bacillus subtilis ATP synthase attenuates ADP-inhibition and decreases fitness in mixed cultures.. Biochem Biophys Res Commun 509(1):102-107 PMID: 30580998
- 8. Bai B et al.. 2018. Identification of candidate genes and long non-coding RNAs associated with the effect of ATP5J in colorectal cancer.. Int J Oncol 52(4):1129-1138 PMID: 29484395
