GO:0106162 mRNA cytidine N-acetyltransferase activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0106162 describes the enzymatic activity that transfers an acetyl group from acetyl-CoA onto cytidine residues within mRNA, forming N4-acetylcytidine (ac4C).
• NAT10 is the principal enzyme responsible for this mRNA cytidine N-acetyltransferase activity in human cells.
• ac4C modification introduced by this activity enhances mRNA stability and translational efficiency, influencing diverse cellular processes.
• Dysregulation of this activity is implicated in cardiac remodeling, chemoresistance, adipogenesis, neuropathic pain, and antiviral immunity.
• Studying GO:0106162 requires integrating RNA modification detection, transcriptomic profiling, and CRISPR-based genetic models.
• EDITGENE provides CRISPR knockout, point-mutation, knock-in, overexpression, and library screening services to dissect the function of this activity in disease models.
Description
GO:0106162, mRNA cytidine N-acetyltransferase activity, is a molecular function that catalyzes the acetylation of cytidine residues within messenger RNA (mRNA) to generate N4-acetylcytidine (ac4C). This enzymatic reaction consumes acetyl-CoA and ATP and produces ADP, CoA, phosphate, and an N4-acetylcytidine-modified mRNA. The discovery of this activity has expanded the field of epitranscriptomics beyond methylation, revealing a new layer of post-transcriptional regulation that affects mRNA fate and cellular physiology. Researchers are increasingly interested in GO:0106162 because it directly links metabolic cofactors to gene expression control and because its dysregulation is associated with human diseases ranging from cancer to cardiac disorders. Understanding this activity at the molecular, cellular, and organismal levels requires combining biochemical assays, transcriptome-wide mapping, and CRISPR-based genetic models.
mRNA cytidine N-acetyltransferase activity At A Glance
| GO ID | GO:0106162 |
|---|---|
| GO term | mRNA cytidine N-acetyltransferase activity |
| Ontology | molecular_function |
| Synonym | mRNA N-acetyltransferase activity |
| Definition | Catalysis of the reaction: a cytidine in mRNA + acetyl-CoA + ATP + H2O = ADP + an N(4)-acetylcytidine in mRNA + CoA + H+ + phosphate. |
| Major function | Introduces N4-acetylcytidine (ac4C) into mRNA, influencing mRNA stability and translation. |
| Representative enzyme | NAT10 (N-acetyltransferase 10) is the primary enzyme known to carry out this activity. |
| Substrates | Cytidine in mRNA, acetyl-CoA, ATP, and water. |
| Products | N4-acetylcytidine in mRNA, ADP, CoA, phosphate, and H+. |
| Associated processes | mRNA stabilization, translational regulation, cellular stress responses, and immune cell expansion. |
What Is GO:0106162?
In our own words, GO:0106162 refers to the catalytic activity that adds an acetyl group to the N4 position of cytidine nucleotides within mRNA molecules. This reaction uses acetyl-CoA as the acetyl donor and ATP as an energy source, releasing ADP, CoA, phosphate, and a proton as byproducts. The resulting modified base, N4-acetylcytidine (ac4C), is an epitranscriptomic mark that can alter mRNA structure, stability, and translation. This activity is distinct from other RNA-modifying enzymes because it specifically targets cytidine in mRNA context and requires both acetyl-CoA and ATP for catalysis.
Why Is mRNA cytidine N-acetyltransferase activity Important in Cell Biology?
GO:0106162 is important because it represents a direct mechanistic link between cellular metabolism and post-transcriptional gene regulation. By depositing ac4C on mRNA, this activity can modulate the stability and translation of specific transcripts, thereby affecting cell proliferation, differentiation, and stress responses. Dysregulation of this activity has been observed in multiple disease contexts, including cancer chemoresistance, cardiac remodeling, neuropathic pain, and impaired antiviral immunity. Therefore, understanding GO:0106162 offers opportunities for therapeutic intervention and biomarker development.
• Regulates mRNA stability and translation efficiency through ac4C modification.
• Controls T cell expansion and antiviral immunity, highlighting a role in immune responses.
• Contributes to cardiac remodeling and fibrosis after myocardial infarction.
• Drives cisplatin chemoresistance in bladder cancer by enhancing DNA repair.
• Promotes adipogenesis via ac4C modification of KLF9 mRNA.
• Facilitates perineural invasion in pancreatic ductal adenocarcinoma through ITGB5 mRNA modification.
• Participates in neutrophil pyroptosis during sepsis by acetylating ULK1 RNA.
• Involved in neuropathic pain by stabilizing SYT9 expression in sensory neurons.
• Provides a target for CRISPR-based functional studies and drug discovery.
• Links acetyl-CoA metabolism to gene expression control, bridging metabolism and epitranscriptomics.
What Happens During mRNA cytidine N-acetyltransferase activity?
Substrate recognition and binding
In simple terms: The enzyme first finds and grabs the mRNA and the acetyl-CoA molecule.
The enzyme, typically NAT10, recognizes specific cytidine residues within mRNA transcripts. This recognition is thought to depend on sequence context and RNA secondary structure, although the exact determinants are still being elucidated. Acetyl-CoA binds to the enzyme's active site, positioning the acetyl group for transfer.
Catalytic transfer of acetyl group
In simple terms: The enzyme snips the acetyl group off acetyl-CoA and attaches it to the cytidine on the mRNA.
In an ATP-dependent manner, the acetyl group from acetyl-CoA is transferred to the N4 position of the target cytidine, forming N4-acetylcytidine (ac4C) in the mRNA. This reaction releases CoA, ADP, phosphate, and a proton as byproducts. The ATP requirement suggests that the enzyme may use energy to drive a conformational change or to activate the substrate.
Post-catalytic mRNA fate
In simple terms: Once modified, the mRNA's behavior changes, often becoming more stable or better translated.
The ac4C mark introduced by this activity can enhance mRNA stability and translational efficiency. For example, ac4C on specific transcripts has been shown to promote their translation and increase protein output. This modification can also affect mRNA localization and interaction with RNA-binding proteins.
Regulation of enzyme activity
In simple terms: The enzyme's activity can be turned up or down by cellular signals.
The activity of NAT10, the main enzyme for GO:0106162, is regulated at multiple levels, including protein expression, post-translational modifications, and interaction with partner proteins. Cellular stress, immune signals, and metabolic status can influence its function. For instance, in T cells, NAT10 activity is required for expansion and antiviral immunity, suggesting tight regulation during immune responses.
Key Genes Involved in GO:0106162 mRNA cytidine N-acetyltransferase activity
The following genes and proteins are central to the function, regulation, and study of GO:0106162.
| Gene | Major Role | Research Relevance |
|---|---|---|
| NAT10 | Primary enzyme catalyzing mRNA cytidine N-acetyltransferase activity | Knockout and overexpression models reveal its role in cardiac remodeling, immunity, and cancer |
| KLF9 | Transcription factor whose mRNA is modified by ac4C | ac4C modification of KLF9 mRNA promotes adipogenesis |
| ITGB5 | Integrin subunit whose mRNA is ac4C-modified | Modification enhances perineural invasion in pancreatic cancer |
| ULK1 | Kinase involved in autophagy, its RNA is acetylated | Acetylation activates STING pathway in sepsis |
| SYT9 | Synaptotagmin involved in neurotransmitter release | ac4C stabilization of SYT9 mRNA contributes to neuropathic pain |
| Amotl1 | Angiomotin-like protein involved in cell polarity | ac4C modification of Amotl1 mRNA drives cardiac fibrosis |
| STING | Immune adaptor protein in cytosolic DNA sensing | Activated downstream of ULK1 RNA acetylation in sepsis |
| TP53 | Tumor suppressor, potential target of ac4C regulation | NAT10-mediated ac4C affects DNA repair and chemoresistance |
| CD4 | T cell co-receptor, marker of T cell expansion | NAT10 activity is required for T cell expansion and antiviral immunity |
| CD8 | Cytotoxic T cell marker | ac4C modification supports effector T cell functions |
| MYC | Oncogene, potential downstream target of ac4C | NAT10 drives chemoresistance possibly via MYC-related pathways |
| ACTB | Housekeeping gene, often used as control | Used as internal control in ac4C studies |
| GAPDH | Glycolytic enzyme, common control | Reference gene in adipogenesis studies |
| IL-6 | Pro-inflammatory cytokine | Induced in sepsis models involving NAT10 |
| TNF-alpha | Pro-inflammatory cytokine | Elevated in sepsis and neuropathic pain models |
| BDNF | Neurotrophic factor | Potential downstream of SYT9 in pain pathways |
| COL1A1 | Collagen type I alpha 1, fibrosis marker | Upregulated in cardiac fibrosis driven by Amotl1 ac4C |
| ACTA2 | Alpha smooth muscle actin, myofibroblast marker | Used to assess cardiac fibrosis |
How Is mRNA cytidine N-acetyltransferase activity Regulated?
The activity of mRNA cytidine N-acetyltransferase is regulated at multiple levels. NAT10 expression can be induced by cellular stress and immune signals. In T cells, NAT10 activity is essential for expansion and antiviral immunity, suggesting regulation by T cell receptor signaling and cytokines. In sepsis, NAT10-mediated acetylation of ULK1 RNA activates the STING pathway, indicating a role in innate immune sensing. Additionally, the availability of acetyl-CoA, a central metabolite, directly influences the reaction rate, linking metabolic status to epitranscriptomic regulation. Post-translational modifications of NAT10, such as phosphorylation and acetylation, may also modulate its enzymatic activity, although specific sites and regulators require further study.
mRNA cytidine N-acetyltransferase activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| NAT10 | Bladder cancer chemoresistance | Knockout bladder cancer cell lines, xenograft models |
| NAT10 | Cardiac remodeling and fibrosis | Knockout mice, myocardial infarction models |
| NAT10 | T cell expansion and antiviral immunity | Conditional knockout mice, viral infection models |
| NAT10 | Sepsis and neutrophil pyroptosis | Knockout mice, LPS-induced sepsis models |
| NAT10 | Neuropathic pain | Knockdown in sensory neurons, nerve injury models |
Cancer and Chemoresistance
NAT10-mediated ac4C modification is implicated in cancer progression and chemoresistance. In bladder cancer, NAT10 drives cisplatin resistance by enhancing ac4C-associated DNA repair pathways. In pancreatic ductal adenocarcinoma, ac4C modification of ITGB5 mRNA promotes perineural invasion, a key step in tumor spread. These findings suggest that targeting GO:0106162 could sensitize tumors to chemotherapy and reduce metastasis.
Cardiovascular Disease
In cardiac remodeling after myocardial infarction, NAT10 is involved in ac4C-mediated transcriptomic regulation, contributing to fibrosis and heart failure. Acetylation of Amotl1 mRNA by NAT10 promotes cardiac fibrotic expansion in mice after myocardial infarction. These studies highlight the potential of modulating this activity to treat cardiac fibrosis.
Immune and Inflammatory Disorders
NAT10-mediated ac4C modification is critical for T cell expansion and antiviral immunity. In sepsis, NAT10 regulates neutrophil pyroptosis by acetylating ULK1 RNA and activating the STING pathway, linking this activity to inflammatory cell death. Dysregulation may contribute to immune pathology, making it a candidate target for immunomodulation.
Neurological and Metabolic Conditions
In neuropathic pain, NAT10 stabilizes SYT9 expression in primary sensory neurons via ac4C modification, suggesting a role in pain sensitization. In adipogenesis, ac4C modification of KLF9 mRNA promotes fat cell differentiation, linking this activity to metabolic regulation. These findings open avenues for treating pain and metabolic disorders.
From mRNA cytidine N-acetyltransferase activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of NAT10 affect mRNA ac4C levels? | CRISPR knockout of NAT10 in cell lines followed by ac4C dot blot or mass spectrometry |
| Does a specific point mutation in NAT10 abolish catalytic activity? | CRISPR point mutation knock-in of catalytic-dead NAT10 |
| Does ac4C modification of a target mRNA affect its stability? | Knock-in of ac4C site mutation in target gene, RNA stability assays |
| Can overexpression of NAT10 drive disease phenotypes? | Overexpression of NAT10 in cell lines or transgenic mice |
| What is the role of NAT10 in immune cell expansion? | Conditional knockout in T cells, adoptive transfer models |
| Can CRISPR library screening identify synthetic lethal partners? | Genome-wide CRISPR knockout library in NAT10-mutant cells |
How to Study the mRNA cytidine N-acetyltransferase activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| ac4C dot blot | Global ac4C levels in RNA | Rapid assessment of changes after NAT10 manipulation |
| Mass spectrometry | Quantitative ac4C modification | Precise measurement of modification stoichiometry |
| ac4C-seq | Transcriptome-wide mapping of ac4C sites | Identifying specific modified transcripts |
| RNA-seq | mRNA abundance changes | Assessing stability effects of ac4C |
| Ribo-seq | Translational efficiency | Determining if ac4C enhances translation |
| CRISPR knockout | Gene function loss | Testing requirement of NAT10 for phenotypes |
| CRISPR point mutation | Catalytic activity | Separating enzymatic from scaffolding functions |
| Immunoprecipitation | Protein-RNA interactions | Identifying ac4C reader proteins |
Detecting ac4C Modifications
To study GO:0106162, researchers use methods such as ac4C dot blot, mass spectrometry, and ac4C-seq to quantify and map N4-acetylcytidine on mRNA. These techniques allow global and transcript-specific measurement of the modification.
Transcriptomic and Translational Profiling
RNA-seq and Ribo-seq can reveal changes in mRNA abundance and translation efficiency upon modulation of this activity. Comparing wild-type and NAT10-knockout cells identifies transcripts whose stability or translation depends on ac4C.
Proteomic and Interaction Studies
Proteomics and immunoprecipitation can identify proteins that interact with NAT10 or recognize ac4C-modified mRNA. These approaches help elucidate the downstream effectors of this activity.
CRISPR-Based Functional Genomics
CRISPR knockout, point mutation, and knock-in models are essential to establish causality between GO:0106162 and specific phenotypes. Library screening can uncover genetic dependencies in cells with altered activity.
How CRISPR Can Be Used to Study GO:0106162 mRNA cytidine N-acetyltransferase activity
Knockout
CRISPR knockout of NAT10 or other genes involved in GO:0106162 allows researchers to assess the loss-of-function consequences on mRNA ac4C levels and downstream phenotypes. For example, NAT10 knockout reduces ac4C and impairs T cell expansion and antiviral immunity.
Point Mutation
Introducing point mutations in the catalytic domain of NAT10 can abolish its acetyltransferase activity without affecting protein levels, helping distinguish enzymatic from non-enzymatic functions. Such models are valuable for dissecting the specific contribution of GO:0106162 to disease.
Knock-in
Knock-in of ac4C site mutations in target mRNAs (e.g., KLF9, ITGB5) can prevent their modification and reveal the functional importance of individual ac4C marks. This approach provides precise mechanistic insights into how this activity affects specific transcripts.
Overexpression
Overexpression of wild-type or mutant NAT10 can drive gain-of-function phenotypes, such as chemoresistance or cardiac fibrosis, and is useful for validating sufficiency. Combining overexpression with ac4C mapping identifies direct targets.
How EDITGENE Supports mRNA cytidine N-acetyltransferase activity Research
Researchers studying mRNA cytidine N-acetyltransferase activity-related genes often need to determine whether a candidate gene is causally involved in a specific phenotype, such as chemoresistance, immune cell expansion, or cardiac fibrosis. Establishing causality requires precise genetic manipulation, which is where EDITGENE's CRISPR services can accelerate discovery.
Contact EDITGENE today to design your custom CRISPR model for mRNA cytidine N-acetyltransferase activity research.
Frequently Asked Questions About mRNA cytidine N-acetyltransferase activity
What is mRNA cytidine N-acetyltransferase activity?
It is the enzymatic activity that adds an acetyl group to cytidine in mRNA, forming N4-acetylcytidine (ac4C), as defined by GO:0106162.
What genes are involved in mRNA cytidine N-acetyltransferase activity?
NAT10 is the primary gene encoding the enzyme for this activity, and target genes include KLF9, ITGB5, ULK1, SYT9, and Amotl1.
What is the role of NAT10 in ac4C modification?
NAT10 catalyzes the transfer of acetyl groups to cytidine residues in mRNA, a key step in ac4C modification.
How does ac4C modification affect mRNA?
ac4C can enhance mRNA stability and translation efficiency, influencing protein expression.
Is mRNA cytidine N-acetyltransferase activity involved in cancer?
Yes, it is implicated in bladder cancer chemoresistance and pancreatic cancer perineural invasion.
What diseases are associated with GO:0106162?
Cardiac remodeling, chemoresistance, adipogenesis, neuropathic pain, sepsis, and impaired antiviral immunity.
How can I study mRNA cytidine N-acetyltransferase activity?
Use ac4C dot blot, mass spectrometry, ac4C-seq, RNA-seq, Ribo-seq, and CRISPR knockout models.
What CRISPR models are available for NAT10 research?
Knockout, point mutation, knock-in, and overexpression models can be generated for NAT10 and its targets.
Does NAT10 have non-enzymatic functions?
Some studies suggest NAT10 may have roles beyond its acetyltransferase activity, but its catalytic function is central to GO:0106162.
Where can I find services to create CRISPR models for ac4C research?
EDITGENE offers custom CRISPR knockout, point mutation, knock-in, overexpression, and library screening services.
Conclusion
GO:0106162, mRNA cytidine N-acetyltransferase activity, represents a critical epitranscriptomic mechanism that regulates mRNA fate and cellular physiology. Its primary enzyme, NAT10, and target transcripts such as KLF9, ITGB5, and ULK1 are implicated in diverse diseases, from cancer to cardiac fibrosis and immune disorders. Continued research using advanced CRISPR models and multi-omics approaches will further illuminate how this activity can be targeted therapeutically.
References
- 1. Shi J et al.. 2023. NAT10 Is Involved in Cardiac Remodeling Through ac4C-Mediated Transcriptomic Regulation.. Circ Res 133(12):989-1002 PMID: 37955115
- 2. Sun L et al.. 2025. A critical role of N(4)-acetylation of cytidine in mRNA by NAT10 in T cell expansion and antiviral immunity.. Nat Immunol 26(4):619-634 PMID: 40045031
- 3. Xie R et al.. 2023. NAT10 Drives Cisplatin Chemoresistance by Enhancing ac4C-Associated DNA Repair in Bladder Cancer.. Cancer Res 83(10):1666-1683 PMID: 36939377
- 4. Wan X et al.. 2025. NAT10-mediated N4-acetylcytidine modification in KLF9 mRNA promotes adipogenesis.. Cell Death Differ 32(9):1613-1629 PMID: 40123006
- 5. Huang L et al.. 2025. N(4)-acetylcytidine modification of ITGB5 mRNA mediated by NAT10 promotes perineural invasion in pancreatic ductal adenocarcinoma.. J Exp Clin Cancer Res 44(1):103 PMID: 40119353
- 6. Zhang H et al.. 2022. NAT10 regulates neutrophil pyroptosis in sepsis via acetylating ULK1 RNA and activating STING pathway.. Commun Biol 5(1):916 PMID: 36068299
- 7. Zhang M et al.. 2023. The Cytidine N-Acetyltransferase NAT10 Participates in Peripheral Nerve Injury-Induced Neuropathic Pain by Stabilizing SYT9 Expression in Primary Sensory Neurons.. J Neurosci 43(17):3009-3027 PMID: 36898834
- 8. Wang XX et al.. 2024. Acetylcytidine modification of Amotl1 by N-acetyltransferase 10 contributes to cardiac fibrotic expansion in mice after myocardial infarction.. Acta Pharmacol Sin 45(7):1425-1437 PMID: 38839936
