GO:0140815 NAD+-protein-histidine ADP-ribosyltransferase activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0140815 describes the enzymatic transfer of an ADP-ribose moiety from NAD+ onto a histidine residue of a target protein, releasing nicotinamide and forming Nt-(ADP-D-ribosyl)-L-histidyl-[protein].
• This activity is a mono-ADP-ribosylation reaction, distinct from poly-ADP-ribosylation, and is catalyzed by members of the ADP-ribosyltransferase (ART) family, including PARP7, PARP15, TIPARP, and bacterial toxins.
• Histidine-specific ADP-ribosylation regulates protein stability, immune signaling, and host-pathogen interactions, with PARP7 and AHR being marked for degradation upon ubiquitin pathway blockade.
• Bacterial effectors such as Clostridium difficile toxins and Pseudomonas ExoY exploit this activity to disrupt host cellular processes, including Rho GTPase signaling and cGMP signaling.
• The reaction can be studied using NAD+ analogs, biocatalytic methods, and inhibitors that target ART domains, enabling drug discovery against pertussis toxin and other pathogens.
• CRISPR-based knockout, point-mutation, knock-in, and overexpression models are essential to dissect the causal roles of ART enzymes in disease and immunity.
Description
NAD+-protein-histidine ADP-ribosyltransferase activity (GO:0140815) is a molecular function that catalyzes the transfer of an ADP-ribose unit from NAD+ to a histidine residue on a protein substrate, producing nicotinamide and an Nt-(ADP-D-ribosyl)-L-histidyl-[protein] adduct. This modification is a form of mono-ADP-ribosylation, a reversible post-translational modification that regulates protein function, stability, and interactions. The reaction is mediated by enzymes containing an ADP-ribosyltransferase (ART) domain, which can be found in eukaryotic proteins such as PARP7, PARP15, and TIPARP, as well as in bacterial toxins like Clostridium difficile toxins and Pseudomonas aeruginosa ExoY. Researchers study this activity because it plays critical roles in immune defense, cell signaling, and host-pathogen interactions, and its dysregulation is linked to cancer, infectious diseases, and inflammatory disorders.
NAD+-protein-histidine ADP-ribosyltransferase activity At A Glance
| GO ID | GO:0140815 |
|---|---|
| GO term | NAD+-protein-histidine ADP-ribosyltransferase activity |
| Ontology | molecular_function |
| Synonym | None |
| Definition | Catalysis of the reaction: L-histidyl-[protein] + NAD+ = H+ + Nt-(ADP-D-ribosyl)-L-histidyl-[protein] + nicotinamide. |
| Major function | Mono-ADP-ribosylation of histidine residues on target proteins, regulating their activity, stability, and interactions. |
| Enzyme family | ADP-ribosyltransferase (ART) domain-containing proteins, including PARPs and bacterial toxins. |
| Substrates | NAD+ and L-histidyl-[protein]. |
| Products | Nicotinamide, H+, and Nt-(ADP-D-ribosyl)-L-histidyl-[protein]. |
| Cellular context | Cytoplasm, nucleus, and extracellular space depending on the enzyme. |
What Is GO:0140815?
GO:0140815 defines the catalytic activity of an enzyme that transfers the ADP-ribose moiety of NAD+ to the nitrogen atom of a histidine residue in a protein, forming an Nt-(ADP-D-ribosyl)-L-histidyl-[protein] linkage and releasing nicotinamide and a proton. This is a mono-ADP-ribosylation event, as opposed to poly-ADP-ribosylation, and it modifies the target protein's chemical properties, often affecting its stability, localization, or interaction partners.
Why Is NAD+-protein-histidine ADP-ribosyltransferase activity Important in Cell Biology?
NAD+-protein-histidine ADP-ribosyltransferase activity is important because it is a key mechanism for post-translational regulation of protein function in both physiological and pathological contexts. It enables rapid and reversible modification of target proteins in response to cellular stress, immune signals, and infection, and it is exploited by bacterial pathogens to subvert host defenses. Understanding this activity is therefore critical for developing therapeutics against infectious diseases, cancer, and inflammatory conditions.
• Regulates protein stability and degradation, as shown for PARP7 and AHR upon ubiquitin pathway blockade.
• Mediates host-pathogen interactions by bacterial toxins such as Clostridium difficile toxins and Pseudomonas ExoY.
• Controls immune signaling and antiviral defense through mRNA ADP-ribosylation.
• Serves as a target for drug discovery, with compounds inhibiting pertussis toxin ART activity identified.
• Involved in cGMP signaling during Pseudomonas aeruginosa infection.
• Modulates transcription factor activity, including AHR and TIPARP.
• Provides a mechanism for NAD+ analog-based biocatalysis and tool development.
• Dysregulation is linked to cancer, neurodegeneration, and inflammatory diseases.
• Enables precise CRISPR-based modeling of gene function in disease research.
• Offers opportunities for targeted therapies using ART domain inhibitors.
What Happens During NAD+-protein-histidine ADP-ribosyltransferase activity?
Substrate Binding and NAD+ Recognition
In simple terms: The enzyme grabs NAD+ and the target protein to start the modification.
The ART domain of the enzyme binds NAD+ and a target protein containing a histidine residue. Structural studies of PARP15 show that dimerization of the ART domain regulates this binding and catalytic activity. Bacterial toxins like Clostridium difficile toxin B also recognize host proteins such as Rho GTPases as substrates.
Catalysis and ADP-Ribose Transfer
In simple terms: The enzyme cuts NAD+ and attaches the ADP-ribose part onto the histidine.
The catalytic reaction cleaves the glycosidic bond of NAD+, releasing nicotinamide and transferring the ADP-ribose moiety to the nitrogen of the histidine residue, forming Nt-(ADP-D-ribosyl)-L-histidyl-[protein]. This mono-ADP-ribosylation is distinct from poly-ADP-ribosylation and can be reversed by cellular enzymes.
Post-Modification Effects on Target Proteins
In simple terms: The added ADP-ribose tag changes how the target protein behaves.
ADP-ribosylation can alter protein stability, localization, and interactions. For example, PARP7 and AHR are marked for degradation following ADP-ribosylation when the ubiquitin pathway is blocked. In antiviral defense, mRNA ADP-ribosylation blocks translation.
Pathogen Exploitation of the Activity
In simple terms: Some bacteria use this reaction to attack host cells.
Bacterial effectors such as Pseudomonas ExoY and Clostridium difficile toxins deliver ART activity into host cells, disrupting signaling pathways like Rho GTPase and cGMP signaling. Inhibitors of pertussis toxin ART activity have been discovered, highlighting therapeutic potential.
Key Genes Involved in GO:0140815 NAD+-protein-histidine ADP-ribosyltransferase activity
The following genes encode enzymes or substrates directly implicated in NAD+-protein-histidine ADP-ribosyltransferase activity, based on published literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| PARP7 | Mono-ADP-ribosyltransferase that marks proteins for degradation | Target for cancer and immune regulation |
| PARP15 | ART domain-containing enzyme regulated by dimerization | Model for studying ART domain regulation |
| TIPARP | Mono-ADP-ribosyltransferase involved in AHR signaling | Methods to study TCDD-inducible activity |
| AHR | Transcription factor regulated by ADP-ribosylation | Degradation upon ubiquitin blockade |
| ExoY | Pseudomonas aeruginosa effector with ART activity | Interplay with cGMP signaling |
| TcdB | Clostridium difficile toxin with ART activity | Targets Rho GTPases |
| Pertussis toxin | Bacterial ART that modifies host proteins | Inhibitor discovery |
| Rho GTPases | Substrates of bacterial ART toxins | Cytoskeletal disruption |
| cGMP | Second messenger affected by ExoY | Infection signaling |
| NAD+ | Cofactor for ADP-ribosylation | Biocatalysis of analogs |
| Histidine | Target residue for modification | Defines GO:0140815 specificity |
| mRNA | Target of antiviral ADP-ribosylation | Translation blockade |
| Ubiquitin | Pathway interplay with ADP-ribosylation | Degradation of PARP7 and AHR |
| ART domain | Catalytic module | Dimerization regulation |
| Nicotinamide | Product of reaction | Biochemical assay |
| ADP-ribose | Transferred moiety | Post-translational modification |
How Is NAD+-protein-histidine ADP-ribosyltransferase activity Regulated?
The activity of NAD+-protein-histidine ADP-ribosyltransferases is regulated at multiple levels. Dimerization of the ART domain in PARP15 controls catalytic activity. The ubiquitin pathway influences the stability of ADP-ribosylated proteins such as PARP7 and AHR. Bacterial effectors like ExoY are activated by host factors and modulate cGMP signaling. Additionally, NAD+ availability and cellular stress can impact the overall rate of ADP-ribosylation.
NAD+-protein-histidine ADP-ribosyltransferase activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| PARP7 | Cancer, immune regulation | Knockout and overexpression cell lines |
| ExoY | Pseudomonas aeruginosa infection | Point mutation of catalytic residues |
| TcdB | Clostridium difficile infection | Knock-in of toxin in host cells |
| Pertussis toxin | Whooping cough | Inhibitor screening with ART mutants |
| TIPARP | AHR signaling, toxicity | Knockout models for ADP-ribosylation |
Cancer and Immune Evasion
ADP-ribosylation by PARP7 and other ART enzymes can mark proteins for degradation, affecting tumor suppressor pathways and immune responses. Targeting this activity may enhance cancer immunotherapy.
Bacterial Infections
Clostridium difficile toxins and Pseudomonas ExoY use ADP-ribosyltransferase activity to disrupt host cell signaling, leading to disease pathology. Inhibitors of pertussis toxin ART activity show promise as anti-infectives.
Antiviral Defense
An mRNA ADP-ribosyltransferase blocks translation as an antiviral mechanism, highlighting the role of this activity in innate immunity.
From NAD+-protein-histidine ADP-ribosyltransferase activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of ART activity affect protein stability? | Knockout of PARP7 or PARP15 |
| Which histidine residue is modified? | Point mutation of target histidine to alanine |
| Can a disease-associated mutation alter activity? | Knock-in of patient-derived mutations |
| Where does the enzyme localize? | Tagged knock-in with fluorescent protein |
| Does overexpression drive transformation? | Overexpression of ART enzymes in cell lines |
| Can inhibitors block bacterial ART? | Bacterial toxin point mutants and inhibitor assays |
How to Study the NAD+-protein-histidine ADP-ribosyltransferase activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| NAD+ analog biocatalysis | ADP-ribose transfer efficiency | Enzyme kinetics |
| Mass spectrometry | ADP-ribosylated histidine sites | Site mapping |
| CRISPR knockout screens | Gene essentiality for ADP-ribosylation | Pathogen-host interaction |
| Inhibitor assays | ART activity inhibition | Drug discovery |
| Ribo-seq | Translation changes upon ADP-ribosylation | Antiviral defense |
| cGMP imaging | Second messenger levels | Pseudomonas infection |
| Dimerization assays | ART domain oligomerization | PARP15 regulation |
| Ubiquitin pathway blockade | Protein degradation of ADP-ribosylated targets | PARP7 and AHR stability |
Biochemical Assays for ADP-Ribosylation
In vitro assays using radiolabeled NAD+ or NAD+ analogs can measure the transfer of ADP-ribose to histidine residues, as demonstrated for TIPARP and PARP15.
Proteomics and Mass Spectrometry
Mass spectrometry can identify ADP-ribosylated histidine sites on target proteins, enabling mapping of modification sites and quantification.
CRISPR Screens and Functional Genomics
Genome-wide CRISPR knockout screens can identify genes required for ADP-ribosylation-dependent phenotypes, such as immune evasion or toxin sensitivity.
Imaging and Cellular Localization
Fluorescent tagging of ART enzymes or substrates allows visualization of localization and dynamics during infection or stress.
How CRISPR Can Be Used to Study GO:0140815 NAD+-protein-histidine ADP-ribosyltransferase activity
Knockout
CRISPR knockout of ART genes such as PARP7 or PARP15 can reveal their roles in protein stability, immune signaling, and disease progression.
Point Mutation
Introducing point mutations in the catalytic histidine or NAD+ binding pocket of ART enzymes can abolish activity and test causality in cellular models.
Knock-in
Knock-in of tagged or disease-associated variants allows tracking of ADP-ribosylation dynamics and localization in live cells.
Overexpression
Overexpression of ART enzymes or bacterial effectors can mimic pathological states and identify downstream signaling changes.
How EDITGENE Supports NAD+-protein-histidine ADP-ribosyltransferase activity Research
Researchers studying NAD+-protein-histidine ADP-ribosyltransferase 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 accelerate this discovery.
Contact EDITGENE today to design your custom CRISPR model for NAD+-protein-histidine ADP-ribosyltransferase activity research.
Frequently Asked Questions About NAD+-protein-histidine ADP-ribosyltransferase activity
What is NAD+-protein-histidine ADP-ribosyltransferase activity?
It is the enzymatic transfer of ADP-ribose from NAD+ to a histidine residue on a protein, forming a mono-ADP-ribosylated product.
What genes are involved in NAD+-protein-histidine ADP-ribosyltransferase activity?
Genes include PARP7, PARP15, TIPARP, and bacterial effectors like ExoY and TcdB.
What is the GO ID for NAD+-protein-histidine ADP-ribosyltransferase activity?
The GO ID is GO:0140815.
How is histidine ADP-ribosylation different from poly-ADP-ribosylation?
Histidine ADP-ribosylation adds a single ADP-ribose unit, while poly-ADP-ribosylation adds chains.
Which diseases are linked to this activity?
It is linked to cancer, bacterial infections, and antiviral defense.
What methods are used to study this activity?
Methods include mass spectrometry, biochemical assays, CRISPR screens, and imaging.
Can CRISPR be used to study NAD+-protein-histidine ADP-ribosyltransferase activity?
Yes, CRISPR knockout, point mutation, knock-in, and overexpression models are widely used.
What are the substrates of this enzyme?
The substrates are NAD+ and a protein containing a histidine residue.
What are the products of the reaction?
The products are nicotinamide, H+, and Nt-(ADP-D-ribosyl)-L-histidyl-[protein].
How is this activity regulated?
It is regulated by ART domain dimerization, ubiquitin pathway interplay, and NAD+ availability.
Conclusion
NAD+-protein-histidine ADP-ribosyltransferase activity (GO:0140815) is a fundamental enzymatic function that controls protein fate and host-pathogen interactions through mono-ADP-ribosylation. Its roles in cancer, infection, and immunity make it a high-value target for therapeutic development. CRISPR-based models and biochemical tools are essential to unravel its mechanisms and translate findings into clinical applications.
References
- 1. Sakari M et al.. 2025. ADP-ribosyltransferase-based biocatalysis of nonhydrolyzable NAD+ analogs.. J Biol Chem 301(1):108106 PMID: 39706271
- 2. Aktories K et al.. 2017. Clostridium difficile Toxin Biology.. Annu Rev Microbiol 71:281-307 PMID: 28657883
- 3. Gorelik A et al.. 2026. Ubiquitin pathway blockade reveals endogenous ADP-ribosylation marking PARP7 and AHR for degradation.. EMBO J 45(1):261-277 PMID: 41326691
- 4. Ebenwaldner C et al.. 2025. Regulation of ADP-ribosyltransferase activity by ART domain dimerization in PARP15.. Nat Commun 16(1):9567 PMID: 41162413
- 5. Vassallo CN et al.. 2024. Anti-viral defence by an mRNA ADP-ribosyltransferase that blocks translation.. Nature 636(8041):190-197 PMID: 39443800
- 6. Hutin D et al.. 2018. Methods to Study TCDD-Inducible Poly-ADP-Ribose Polymerase (TIPARP) Mono-ADP-Ribosyltransferase Activity.. Methods Mol Biol 1813:109-124 PMID: 30097864
- 7. Ashok Y et al.. 2020. Discovery of Compounds Inhibiting the ADP-Ribosyltransferase Activity of Pertussis Toxin.. ACS Infect Dis 6(4):588-602 PMID: 31899865
- 8. Deruelle V et al.. 2025. Interplay between T3SS effectors, ExoY activation, and cGMP signaling in Pseudomonas aeruginosa infection.. Nat Commun 17(1):69 PMID: 41330935
