GO:0061733 protein-lysine-acetyltransferase activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0061733 protein-lysine-acetyltransferase activity catalyzes the transfer of an acetyl group from acetyl-CoA to the epsilon-amino group of a lysine residue on a protein, releasing CoA and H+.
• This activity is central to epigenetic regulation, metabolism, and signal transduction, with CBP/p300 being a major human acetyltransferase frequently dysregulated in cancer.
• Allosteric communication pathways regulate the catalytic efficiency of protein lysine acetyltransferases, making them attractive drug targets.
• In Mycobacterium tuberculosis, cyclic AMP-dependent protein lysine acetylation controls fatty acid and propionate metabolism, linking this activity to bacterial pathogenesis.
• Histone lactylation, a recently discovered modification, is driven by protein lysine acetyltransferase activity and contributes to cancer progression.
• Small-molecule inhibitors of p300 histone acetyltransferase, such as quinoline-based compounds, induce apoptosis in leukemia cells, validating this activity as a therapeutic target.
Description
Protein-lysine-acetyltransferase activity (GO:0061733) is a fundamental enzymatic function that transfers an acetyl group from acetyl-CoA to the side-chain amino group of lysine residues in proteins, producing N6-acetyl-L-lysyl-[protein], CoA, and a proton. This post-translational modification is reversible and dynamically regulated, influencing protein stability, localization, interactions, and enzymatic activity. The reaction is conserved from bacteria to humans and plays a pivotal role in diverse cellular processes, including transcription, metabolism, and stress responses. In eukaryotes, histone acetyltransferases (HATs) such as CBP/p300 acetylate histone tails to modulate chromatin structure and gene expression, while non-histone substrates are involved in cell cycle control and apoptosis. In prokaryotes, protein lysine acetylation regulates metabolic enzymes and virulence factors, as demonstrated in Mycobacterium tuberculosis where cyclic AMP signaling controls acetylation of metabolic proteins. The importance of GO:0061733 extends to human disease: dysregulation of acetyltransferases is implicated in cancer, neurodegeneration, and metabolic disorders, making these enzymes promising therapeutic targets. Understanding the molecular mechanisms, key genes, and regulatory networks of protein-lysine-acetyltransferase activity is therefore essential for both basic research and drug development.
protein-lysine-acetyltransferase activity At A Glance
| GO ID | GO:0061733 |
|---|---|
| GO term | protein-lysine-acetyltransferase activity |
| Ontology | molecular_function |
| Synonym | peptide-lysine acetyltransferase activity; protein acetyltransferase activity; protein-lysine-N-acetylase activity |
| Major function | Catalyzes acetyl transfer from acetyl-CoA to lysine residues on proteins, forming N6-acetyl-L-lysine |
| Reaction | L-lysyl-[protein] + acetyl-CoA = N6-acetyl-L-lysyl-[protein] + CoA + H+ |
| Cofactor | Acetyl-CoA as acetyl donor |
| Subcellular location | Nucleus, cytoplasm, mitochondria (varies by enzyme) |
| Representative enzymes | CBP/p300, GCN5, MYST family, bacterial Pat |
What Is GO:0061733?
Protein-lysine-acetyltransferase activity (GO:0061733) is defined as the catalysis of the reaction: L-lysyl-[protein] + acetyl-CoA = N6-acetyl-L-lysyl-[protein] + CoA + H+. In other words, it is an enzymatic activity that covalently attaches an acetyl group to a lysine residue on a target protein, using acetyl-coenzyme A as the acetyl donor. This activity is synonymous with peptide-lysine acetyltransferase, protein acetyltransferase, and related terms. It is a molecular function that enables the post-translational modification known as lysine acetylation, which can alter protein function, interactions, and stability.
Why Is protein-lysine-acetyltransferase activity Important in Cell Biology?
Protein-lysine-acetyltransferase activity is crucial because it governs a vast array of biological processes through reversible lysine acetylation. This modification affects chromatin remodeling, gene transcription, DNA repair, cell cycle progression, and metabolic flux. In cancer, aberrant acetylation driven by enzymes such as CBP/p300 promotes oncogenic gene expression and tumor growth, making these enzymes high-priority drug targets. In infectious diseases, bacterial acetyltransferases regulate virulence and metabolic adaptation, as shown for Mycobacterium tuberculosis. Moreover, emerging roles in histone lactylation link this activity to cancer metabolism and immune responses. Thus, understanding GO:0061733 is essential for deciphering disease mechanisms and developing targeted therapies.
• Regulates gene expression by acetylating histones and non-histone proteins, impacting chromatin structure and transcription.
• Controls metabolic pathways through acetylation of enzymes involved in fatty acid and propionate metabolism in bacteria.
• Drives cancer progression via CBP/p300-mediated acetylation of oncogenes and tumor suppressors.
• Modulates apoptosis; inhibition of p300 HAT activity induces cell death in leukemia cells.
• Links cellular metabolism to epigenetic regulation through histone lactylation.
• Plays a role in bacterial pathogenesis and siderophore biosynthesis in Mycobacterium tuberculosis.
• Represents a druggable target for small-molecule inhibitors in cancer and infectious diseases.
• Involved in mitochondrial protein acetylation, influencing fat storage and energy metabolism.
• Allosteric regulation of acetyltransferases offers opportunities for selective modulation.
• Essential for post-translational control of protein function in both prokaryotes and eukaryotes.
What Happens During protein-lysine-acetyltransferase activity?
Substrate Recognition and Binding
In simple terms: The enzyme first grabs the target protein and the acetyl donor molecule.
Protein lysine acetyltransferases (KATs) recognize specific lysine residues on target proteins through structural motifs that confer substrate specificity. For example, CBP/p300 contains a bromodomain that binds acetylated lysines and a HAT domain that engages the substrate. Allosteric communication pathways within the enzyme facilitate conformational changes required for substrate binding and catalysis. In Mycobacterium tuberculosis, cyclic AMP binding to the acetyltransferase Pat regulates its activity and substrate selection.
Acetyl Group Transfer
In simple terms: The enzyme moves an acetyl group from acetyl-CoA onto the lysine, like attaching a small tag.
The catalytic mechanism involves a direct nucleophilic attack of the lysine epsilon-amino group on the acetyl-CoA thioester, forming a ternary complex. This results in the transfer of the acetyl group to the lysine, releasing coenzyme A and a proton. The reaction is highly conserved and requires a general base to deprotonate the lysine, often assisted by surrounding residues. In bacteria, this acetylation can modulate enzyme activity, as seen for MbtA in siderophore biosynthesis.
Product Release and Protein Conformational Change
In simple terms: After tagging, the enzyme lets go, and the tagged protein may change shape or function.
Following acetylation, the modified protein undergoes conformational changes that can alter its interactions, localization, or activity. For instance, acetylation of histone tails neutralizes positive charges, loosening chromatin and promoting transcription. In metabolic enzymes, acetylation can inhibit or activate catalytic function, as observed in mycobacterial fatty acid metabolism. The release of CoA and H+ completes the reaction, and the enzyme is ready for another cycle.
Regulation by Allostery and Post-translational Modifications
In simple terms: The enzyme's activity can be turned up or down by other molecules or modifications.
Allosteric communication networks within KATs allow remote regulatory sites to influence catalysis. For example, cyclic AMP binds to mycobacterial Pat and modulates its acetyltransferase activity, affecting downstream metabolic pathways. Additionally, autoacetylation of CBP/p300 regulates its own activity and interaction with partners. These regulatory mechanisms ensure precise control of acetylation in response to cellular signals.
Key Genes Involved in GO:0061733 protein-lysine-acetyltransferase activity
The following genes encode proteins with protein-lysine-acetyltransferase activity or are directly involved in its regulation and downstream effects.
| Gene | Major Role | Research Relevance |
|---|---|---|
| CREBBP | Histone acetyltransferase CBP; acetylates histones and non-histone proteins | Frequently mutated in cancer; target for inhibitors |
| EP300 | Histone acetyltransferase p300; coactivator of transcription | Oncogenic driver; small-molecule inhibitors under development |
| KAT2A | GCN5; histone acetyltransferase in SAGA complex | Role in chromatin remodeling and cancer |
| KAT2B | PCAF; acetylates histones and p53 | Implicated in cell cycle and apoptosis |
| KAT5 | TIP60; acetylates histones and ATM | DNA damage response and cancer |
| KAT6A | MOZ; histone acetyltransferase | Leukemogenesis and developmental disorders |
| KAT7 | MYST2; acetylates histone H4 | Regulation of gene expression and replication |
| KAT8 | MOF; acetylates histone H4K16 | Chromatin structure and cancer |
| NCOA1 | SRC-1; coactivator with intrinsic acetyltransferase activity | Nuclear receptor signaling |
| NCOA3 | SRC-3; acetyltransferase coactivator | Breast cancer and metastasis |
| CLOCK | Circadian acetyltransferase; acetylates histones and non-histone proteins | Metabolic regulation and circadian rhythm |
| Pat | Mycobacterial protein lysine acetyltransferase | Regulates fatty acid and propionate metabolism |
| MbtA | Mycobacterial siderophore biosynthesis enzyme; acetylated by Pat | Virulence and iron acquisition |
| ACAT1 | Mitochondrial acetyl-CoA acetyltransferase | Ketone body metabolism and acetylation |
| SIRT1 | Deacetylase (opposing activity) | Regulates acetylation homeostasis |
| HDAC1 | Histone deacetylase (opposing activity) | Cancer and epigenetic therapy |
| GCN5L2 | Histone acetyltransferase | Transcription and development |
How Is protein-lysine-acetyltransferase activity Regulated?
Protein-lysine-acetyltransferase activity is regulated at multiple levels. Allosteric communication pathways within the enzyme can modulate catalytic efficiency in response to ligand binding. In Mycobacterium tuberculosis, cyclic AMP binds to the acetyltransferase Pat, altering its activity and substrate specificity to control fatty acid and propionate metabolism. Post-translational modifications, including autoacetylation, regulate CBP/p300 activity and its interactions with transcription factors. Additionally, the availability of acetyl-CoA, which is influenced by metabolic state, directly affects acetyltransferase activity. Opposing deacetylases, such as sirtuins and HDACs, maintain dynamic acetylation balance.
protein-lysine-acetyltransferase activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| CREBBP | Cancer (leukemia, solid tumors) | Knockout or point-mutation in cancer cell lines; xenograft models |
| EP300 | Cancer (leukemia, breast cancer) | Overexpression and inhibitor testing in U937 cells |
| Pat (Mycobacterium tuberculosis) | Tuberculosis pathogenesis | Knockout in M. tuberculosis; infection models |
| MbtA | Tuberculosis virulence | Point mutation of acetylation sites; siderophore assays |
| CLOCK | Metabolic syndrome, circadian disorders | Knockout mice; metabolic phenotyping |
Cancer
Dysregulation of protein-lysine-acetyltransferase activity is a hallmark of many cancers. CBP/p300 are frequently mutated or overexpressed in solid tumors and hematological malignancies, where they drive oncogenic transcription programs. Small-molecule inhibitors targeting the HAT domain of p300, such as quinoline-based compounds, induce apoptosis in leukemia cells, highlighting the therapeutic potential of inhibiting this activity. Histone lactylation, a newly recognized modification dependent on acetyltransferase activity, promotes cancer progression by altering gene expression.
Infectious Diseases
In Mycobacterium tuberculosis, protein lysine acetyltransferase activity regulates metabolic pathways essential for survival and virulence. Cyclic AMP-dependent acetylation of metabolic enzymes controls fatty acid and propionate metabolism, and acetylation of MbtA modulates siderophore biosynthesis, affecting iron acquisition. Targeting these bacterial acetyltransferases could provide new strategies against tuberculosis.
Metabolic Disorders
Mitochondrial protein acetylation, mediated by acetyltransferases and opposed by sirtuins, is a key regulator of fat storage and energy metabolism. Chemical and metabolic logic of acetyl-lysine modifications links this activity to obesity and insulin resistance. Dysregulation of acetylation balance contributes to metabolic syndrome.
From protein-lysine-acetyltransferase activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of CBP/p300 acetyltransferase activity affect tumor growth? | CRISPR knockout of CREBBP/EP300 in cancer cell lines and mouse xenografts |
| What is the role of specific acetylation sites on a target protein? | Point mutation (lysine to arginine) knock-in via CRISPR |
| How does acetylation of a metabolic enzyme alter its function? | Knock-in of acetylation-mimetic (lysine to glutamine) or non-acetylatable (lysine to arginine) mutants |
| Can a small molecule inhibit p300 HAT activity in vivo? | Overexpression of EP300 in leukemia cells followed by inhibitor treatment |
| What is the impact of bacterial acetyltransferase on virulence? | Knockout of Pat in Mycobacterium tuberculosis and infection in macrophages |
| How does histone lactylation affect gene expression? | Knock-in of histone mutants or overexpression of lactyltransferases |
How to Study the protein-lysine-acetyltransferase activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Acetyl-lysine proteomics | Global acetylation sites and stoichiometry | Mapping substrates of CBP/p300 |
| In vitro HAT assay | Enzymatic activity and kinetics | Screening small-molecule inhibitors |
| ChIP-seq | Genomic localization of acetylated histones | Studying chromatin regulation |
| CRISPR knockout screen | Gene essentiality and modifiers of acetylation | Identifying synthetic lethal partners |
| Western blot with acetyl-lysine antibodies | Specific protein acetylation levels | Validating target acetylation |
| Metabolic flux analysis | Pathway activity affected by acetylation | Bacterial metabolism studies |
| Surface plasmon resonance | Binding affinity of acetyltransferase to substrates | Allosteric modulation studies |
| X-ray crystallography | Three-dimensional structure of enzyme-substrate complexes | Structure-based drug design |
Proteomics and Acetylome Profiling
Mass spectrometry-based acetylome profiling using acetyl-lysine enrichment allows global identification of acetylation sites and quantification of changes upon perturbation of acetyltransferase activity. This method is essential for mapping substrates and understanding the scope of GO:0061733.
Enzymatic Activity Assays
In vitro acetyltransferase assays using recombinant enzymes and acetyl-CoA measure catalytic activity and kinetics. Fluorescent or radioactive acetyl-CoA donors enable high-throughput screening for inhibitors.
Chromatin Immunoprecipitation (ChIP)
ChIP with antibodies against acetylated histones or acetyltransferases identifies genomic regions where acetylation occurs, linking GO:0061733 to transcriptional regulation.
CRISPR-Based Genetic Screens
Genome-wide CRISPR knockout or activation screens can identify genes that modulate acetyltransferase activity or acetylation-dependent phenotypes, revealing novel regulators and therapeutic targets.
How CRISPR Can Be Used to Study GO:0061733 protein-lysine-acetyltransferase activity
Knockout
CRISPR knockout of genes encoding protein-lysine-acetyltransferases (e.g., CREBBP, EP300) enables loss-of-function studies to determine their role in cell proliferation, differentiation, and disease. Knockout cell lines are valuable for identifying acetylation-dependent phenotypes and validating drug targets.
Point Mutation
Point mutations can be introduced to ablate catalytic activity (e.g., mutation of the catalytic glutamate) or to mimic acetylation (lysine to glutamine) or prevent it (lysine to arginine) on substrate proteins. These models help dissect the specific contribution of individual acetylation sites to protein function and disease.
Knock-in
Knock-in of tagged acetyltransferases (e.g., FLAG, HA) allows for affinity purification and interaction studies. Knock-in of disease-associated mutations (e.g., in CREBBP) creates isogenic models to study altered acetyltransferase activity in cancer or developmental disorders.
Overexpression
CRISPR activation (CRISPRa) or cDNA overexpression of acetyltransferases can model gain-of-function states observed in cancer. Overexpression of EP300 in leukemia cells increases histone acetylation and can be used to test the efficacy of HAT inhibitors.
How EDITGENE Supports protein-lysine-acetyltransferase activity Research
Researchers studying protein-lysine-acetyltransferase activity-related genes often need to determine whether a candidate gene is causally involved in a specific biological process or disease. EDITGENE provides comprehensive CRISPR-based services to create precisely engineered cell models, enabling functional validation of acetyltransferases and their substrates.
Contact EDITGENE today to design your custom CRISPR model for protein-lysine-acetyltransferase activity research.
Frequently Asked Questions About protein-lysine-acetyltransferase activity
What is protein-lysine-acetyltransferase activity?
It is an enzymatic activity (GO:0061733) that transfers an acetyl group from acetyl-CoA to a lysine residue on a protein, forming N6-acetyl-L-lysine and releasing CoA and H+.
What genes are involved in protein-lysine-acetyltransferase activity?
Key genes include CREBBP (CBP), EP300 (p300), KAT2A (GCN5), KAT2B (PCAF), KAT5 (TIP60), and bacterial Pat, among others.
How is protein-lysine-acetyltransferase activity regulated?
It is regulated by allosteric interactions, post-translational modifications such as autoacetylation, and availability of acetyl-CoA, as well as by cyclic AMP in bacteria.
What diseases are associated with protein-lysine-acetyltransferase activity?
Dysregulation is linked to cancer, infectious diseases like tuberculosis, and metabolic disorders.
What is the role of CBP/p300 in cancer?
CBP/p300 are frequently mutated or overexpressed in cancer and drive oncogenic transcription; inhibitors are being developed as anticancer drugs.
How can I study protein-lysine-acetyltransferase activity in the lab?
Common methods include acetylome proteomics, in vitro HAT assays, ChIP-seq, and CRISPR screens.
What is histone lactylation?
It is a recently discovered histone modification driven by acetyltransferase activity that links metabolism to gene expression and cancer progression.
Can CRISPR be used to study acetyltransferases?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models enable functional studies of acetyltransferases and their substrates.
What is the reaction catalyzed by protein-lysine-acetyltransferase?
L-lysyl-[protein] + acetyl-CoA = N6-acetyl-L-lysyl-[protein] + CoA + H+.
Why is protein-lysine-acetyltransferase a drug target?
Because its dysregulation contributes to cancer and infectious diseases, and small-molecule inhibitors have shown efficacy in preclinical models.
Conclusion
Protein-lysine-acetyltransferase activity (GO:0061733) is a central post-translational modification mechanism that regulates diverse cellular processes and is implicated in major human diseases. Understanding its molecular mechanisms, key genes, and regulatory networks is essential for developing targeted therapies. EDITGENE provides comprehensive CRISPR services to facilitate functional studies of acetyltransferases and accelerate drug discovery.
References
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- 2. Rehman AU et al.. 2020. Decoding allosteric communication pathways in protein lysine acetyltransferase.. Int J Biol Macromol 149:70-80 PMID: 31987943
- 3. Jia Z et al.. 2026. From mechanism to targeted therapy: Advances in histone lactylation-driven cancer progression (Review).. Oncol Lett 31(1):28 PMID: 41277911
- 4. Lenoci A et al.. 2014. Quinoline-based p300 histone acetyltransferase inhibitors with pro-apoptotic activity in human leukemia U937 cells.. ChemMedChem 9(3):542-8 PMID: 24504685
- 5. Ghanta S et al.. 2013. Mitochondrial protein acetylation as a cell-intrinsic, evolutionary driver of fat storage: chemical and metabolic logic of acetyl-lysine modifications.. Crit Rev Biochem Mol Biol 48(6):561-74 PMID: 24050258
- 6. Lee HJ et al.. 2012. Cyclic AMP regulation of protein lysine acetylation in Mycobacterium tuberculosis.. Nat Struct Mol Biol 19(8):811-8 PMID: 22773105
- 7. Nambi S et al.. 2013. Cyclic AMP-dependent protein lysine acylation in mycobacteria regulates fatty acid and propionate metabolism.. J Biol Chem 288(20):14114-14124 PMID: 23553634
- 8. Vergnolle O et al.. 2016. Post-translational Acetylation of MbtA Modulates Mycobacterial Siderophore Biosynthesis.. J Biol Chem 291(42):22315-22326 PMID: 27566542
