GO:0140822 NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0140822 describes the enzymatic transfer of ADP-ribose from NAD+ to the glutamate-35 residue of histone H2B, a chromatin modification that can alter gene expression.
• This activity is a molecular function that couples cellular NAD+ metabolism to epigenetic regulation, linking metabolic state to transcriptional output.
• Glutamate ADP-ribosylation is a reversible post-translational modification, and its dysregulation has been implicated in cancer, neurodegeneration, and inflammatory diseases.
• Key proteins involved include histone H2B, NAD+-dependent ADP-ribosyltransferases, and NAD+ biosynthetic enzymes such as NAMPT and NMNAT.
• Studying this activity requires tools such as CRISPR knockout, point-mutation, and knock-in cell models, combined with proteomics and sequencing-based readouts.
• EDITGENE provides custom CRISPR cell models and screening services to dissect the function of GO:0140822 in health and disease.
Description
GO:0140822, NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity, is a molecular function that catalyzes the transfer of ADP-ribose groups from NAD+ to the glutamate-35 residue of histone H2B. This modification, known as histone ADP-ribosylation, represents a direct link between cellular energy metabolism and chromatin regulation. Unlike other histone modifications such as acetylation or methylation, ADP-ribosylation is bulky and carries two negative charges, which can significantly impact chromatin structure and protein-protein interactions. Understanding this activity is crucial because it sits at the intersection of NAD+ biology, epigenetic control, and stress responses. Recent studies have begun to uncover the roles of histone ADP-ribosylation in processes such as DNA repair, transcription, and cell death. The specificity for glutamate-35 of histone H2B suggests a targeted regulatory mechanism that may fine-tune gene expression programs. However, the enzymes responsible for this activity and their physiological relevance remain incompletely understood. This article synthesizes current knowledge on GO:0140822, covering its definition, mechanism, key genes, disease associations, and research methodologies, to provide a comprehensive resource for biomedical researchers.
NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity At A Glance
| GO ID | GO:0140822 |
|---|---|
| GO term | NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity |
| Ontology | molecular_function |
| Synonym | NAD+-histone-glutamate ADP-ribosyltransferase activity (H2B-E35 specific), NAD+-histone H2B-E35 glutamate ADP-ribosyltransferase activity |
| Major function | Transfer of ADP-ribose from NAD+ to glutamate-35 of histone H2B |
| Substrate | NAD+ and histone H2B (glutamate-35) |
| Product | ADP-ribosylated histone H2B and nicotinamide |
| Reversibility | Reversible modification, removed by ADP-ribosylhydrolases |
| Cellular context | Nucleus, chromatin |
What Is GO:0140822?
According to the Gene Ontology, GO:0140822 is defined as the catalysis of the transfer of ADP-ribose groups to the glutamate-35 residue of the N-terminal tail of histone H2B (or an equivalent residue). This activity uses NAD+ as a substrate and releases nicotinamide, attaching the ADP-ribose moiety to the glutamate side chain. It is a molecular function that specifically targets a glutamate residue, distinguishing it from serine or arginine ADP-ribosylation.
Why Is NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity Important in Cell Biology?
GO:0140822 is important because it directly connects cellular NAD+ levels to epigenetic regulation, thereby linking metabolic status to gene expression. This activity can influence chromatin accessibility, transcription factor recruitment, and DNA repair processes. Dysregulation of histone ADP-ribosylation has been associated with cancer, neurodegenerative disorders, and inflammatory conditions, making it a potential therapeutic target. Moreover, understanding this activity can shed light on how cells respond to metabolic stress and maintain genome stability.
• Links NAD+ metabolism to epigenetic regulation.
• Modulates chromatin structure and gene transcription.
• Plays a role in DNA damage response and repair.
• Implicated in cancer development and progression.
• Associated with neurodegenerative diseases.
• Involved in inflammatory and immune responses.
• Potential target for therapeutic intervention.
• Requires advanced CRISPR models for functional studies.
• Cross-talks with other histone modifications.
• Can be studied using proteomics and sequencing approaches.
What Happens During NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity?
Substrate Recognition and Binding
In simple terms: The enzyme finds and binds to histone H2B and NAD+.
The enzymatic reaction begins with the recognition of histone H2B, specifically the N-terminal tail containing glutamate-35, by an ADP-ribosyltransferase enzyme. Concurrently, NAD+ binds to the catalytic domain of the enzyme. This dual substrate binding ensures specificity for both the histone target and the cofactor.
Catalysis and ADP-Ribose Transfer
In simple terms: The enzyme cuts NAD+ and attaches the ADP-ribose part to histone H2B.
Upon binding, the enzyme catalyzes the cleavage of NAD+ into nicotinamide and ADP-ribose. The ADP-ribose moiety is then transferred to the glutamate-35 residue of histone H2B, forming a covalent bond. This reaction is a key step in histone ADP-ribosylation and results in a bulky, negatively charged modification on the histone tail.
Chromatin Remodeling and Functional Consequences
In simple terms: The added ADP-ribose changes how DNA is packaged and read.
The addition of ADP-ribose to histone H2B can alter chromatin structure by affecting nucleosome stability and interactions with other proteins. This modification may recruit chromatin remodelers or transcription factors, thereby influencing gene expression. It can also serve as a signal for DNA repair machinery at sites of damage.
Reversal and Regulation
In simple terms: Other enzymes can remove the ADP-ribose to reverse the modification.
Histone ADP-ribosylation is reversible; enzymes such as ADP-ribosylhydrolases can remove the ADP-ribose group from glutamate-35. This reversibility allows dynamic regulation of the modification in response to cellular signals. The balance between addition and removal is critical for normal cellular function.
Key Genes Involved in GO:0140822 NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity
The following genes and proteins are key players in the regulation and function of NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity.
| Gene | Major Role | Research Relevance |
|---|---|---|
| H2B | Histone substrate for ADP-ribosylation at glutamate-35 | Core target for modification studies |
| NAMPT | Rate-limiting enzyme in NAD+ biosynthesis | Regulates NAD+ availability for the reaction |
| NMNAT | NAD+ synthase | Maintains nuclear NAD+ pools |
| PARP1 | ADP-ribosyltransferase; potential enzyme for histone modification | Candidate writer for H2B ADP-ribosylation |
| PARP2 | ADP-ribosyltransferase | Potential redundant or specific role |
| ARTD1 | ADP-ribosyltransferase | May catalyze the modification |
| ARTD2 | ADP-ribosyltransferase | May catalyze the modification |
| SIRT1 | NAD+-dependent deacetylase | Cross-talk with ADP-ribosylation |
| SIRT6 | NAD+-dependent deacetylase and ADP-ribosyltransferase | Potential dual function |
| MACROD1 | ADP-ribosylhydrolase | Removes ADP-ribose from histones |
| MACROD2 | ADP-ribosylhydrolase | Reverses the modification |
| TARG1 | ADP-ribosylhydrolase | Reverses the modification |
| PARG | Poly(ADP-ribose) glycohydrolase | Degrades poly(ADP-ribose) chains |
| HPF1 | Accessory factor for PARP1 | Modulates ADP-ribosylation specificity |
| NADSYN1 | NAD+ synthetase | Contributes to NAD+ pool |
| NMNAT1 | Nuclear NAD+ synthase | Local NAD+ supply for chromatin reactions |
| NMNAT2 | Cytoplasmic NAD+ synthase | Indirect role via NAD+ homeostasis |
| NMNAT3 | Mitochondrial NAD+ synthase | Indirect role via NAD+ homeostasis |
How Is NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity Regulated?
The activity of NAD+-histone H2BE35 glutamate ADP-ribosyltransferase is regulated by cellular NAD+ levels, which are influenced by biosynthetic enzymes such as NAMPT and NMNAT. Additionally, the expression and activity of ADP-ribosyltransferases and hydrolases are controlled by developmental and stress signals. Post-translational modifications of these enzymes and their interaction partners further modulate the reaction.
NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| H2B | Cancer, neurodegeneration | Point mutation at E35 (E35A) in cell lines |
| PARP1 | Cancer, inflammation | Knockout and overexpression models |
| NAMPT | Metabolic disorders, cancer | Knockout and knock-in models |
| MACROD1 | Neurodegeneration | Knockout and overexpression models |
| SIRT6 | Cancer, aging | Knockout and point mutation models |
Cancer
Dysregulated histone ADP-ribosylation has been observed in various cancers, where it can promote oncogenic transcription programs and genomic instability. For example, increased ADP-ribosylation of histone H2B at glutamate-35 may enhance the expression of genes involved in cell proliferation and survival. Targeting this activity could therefore be a therapeutic strategy.
Neurodegeneration
In neurodegenerative diseases such as Alzheimer's and Parkinson's, altered NAD+ metabolism and histone ADP-ribosylation have been reported. Oxidative stress and DNA damage can trigger excessive ADP-ribosylation, contributing to neuronal dysfunction. Modulating this modification may offer neuroprotective benefits.
Inflammatory Diseases
Histone ADP-ribosylation is involved in the regulation of inflammatory gene expression. In conditions like sepsis and autoimmune disorders, aberrant ADP-ribosylation may exacerbate inflammation. Inhibitors of ADP-ribosyltransferases are being explored for their anti-inflammatory effects.
From NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| What is the role of H2B E35 ADP-ribosylation in transcription? | Point mutation (E35A) knock-in cell line |
| Which enzyme catalyzes this modification? | Knockout of candidate ADP-ribosyltransferases (e.g., PARP1, PARP2) |
| How does NAD+ availability affect the modification? | Knockout or overexpression of NAMPT/NMNAT |
| What are the downstream effects on chromatin? | Tagged knock-in of H2B with epitope tag for ChIP-seq |
| Can we reverse the modification? | Overexpression of ADP-ribosylhydrolases (e.g., MACROD1) |
| What is the kinetic profile of the enzyme? | In vitro assays with recombinant proteins |
How to Study the NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Mass spectrometry | ADP-ribosylated peptides and sites | Identification of H2B E35 modification |
| ChIP-seq | Genomic localization of ADP-ribosylation | Mapping modification across the genome |
| CRISPR knockout | Loss of enzyme function | Determining necessity of candidate enzymes |
| CRISPR knock-in | Tagged or mutant histone H2B | Tracking modification and function |
| NAD+ metabolomics | Cellular NAD+ levels | Linking metabolism to modification |
| In vitro ADP-ribosylation assay | Enzymatic activity | Biochemical characterization of enzymes |
| RNA-seq | Transcriptional changes | Assessing impact on gene expression |
Proteomics and Mass Spectrometry
Mass spectrometry can identify and quantify ADP-ribosylated peptides, including histone H2B E35, providing site-specific information. This approach is essential for validating the modification and studying its dynamics.
Chromatin Immunoprecipitation (ChIP)
ChIP using antibodies against ADP-ribose or tagged histones can map the genomic distribution of the modification. Coupled with sequencing (ChIP-seq), it reveals associations with specific chromatin states.
CRISPR-Cas9 Genome Editing
CRISPR knockout, knock-in, and point mutation models allow functional dissection of the enzymes and histone residues involved. These models are crucial for linking the modification to cellular phenotypes.
NAD+ Metabolomics
Quantification of NAD+ and its metabolites by LC-MS informs on the metabolic state that regulates the activity. This can be combined with genetic perturbations to study causality.
How CRISPR Can Be Used to Study GO:0140822 NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity
Knockout
CRISPR knockout of candidate ADP-ribosyltransferases (e.g., PARP1, PARP2) or NAD+ biosynthetic enzymes (e.g., NAMPT) can abolish or reduce H2B E35 ADP-ribosylation, allowing researchers to identify the responsible enzymes and study downstream effects.
Point Mutation
Introducing a point mutation at the catalytic residue of an ADP-ribosyltransferase or at histone H2B E35 (e.g., E35A) can specifically disable the modification without affecting other functions, providing precise mechanistic insights.
Knock-in
Knock-in of tagged histone H2B (e.g., HA or FLAG) enables affinity purification and ChIP-based mapping of the modified histone, facilitating genome-wide studies.
Overexpression
Overexpression of wild-type or mutant ADP-ribosyltransferases or hydrolases can elevate or reduce modification levels, helping to establish causality and dose-dependent effects.
How EDITGENE Supports NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity Research
Researchers studying NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity-related genes often need to determine whether a candidate gene is causally involved in the modification and its downstream phenotypes. EDITGENE provides a comprehensive suite of CRISPR services to accelerate this research.
Contact EDITGENE today to design your custom CRISPR model for NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity research.
Frequently Asked Questions About NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity
What is NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity?
It is a molecular function that transfers ADP-ribose from NAD+ to the glutamate-35 residue of histone H2B, a modification involved in chromatin regulation.
What genes are involved in NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity?
Key genes include histone H2B, NAD+ biosynthetic enzymes like NAMPT and NMNAT, and ADP-ribosyltransferases such as PARP1 and PARP2.
What is the role of histone H2B E35 ADP-ribosylation in cancer?
It can promote oncogenic transcription and genomic instability, making it a potential therapeutic target.
How can I study NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity?
Using CRISPR knockout, point mutation, and knock-in models combined with proteomics, ChIP-seq, and metabolomics.
Which enzymes catalyze histone H2B E35 ADP-ribosylation?
Candidate enzymes include PARP1, PARP2, and other ADP-ribosyltransferases, though the exact enzyme may be context-dependent.
Is histone ADP-ribosylation reversible?
Yes, it is reversed by ADP-ribosylhydrolases such as MACROD1, MACROD2, and TARG1.
What diseases are associated with dysregulated histone ADP-ribosylation?
Cancer, neurodegenerative diseases, and inflammatory conditions.
How does NAD+ metabolism affect histone ADP-ribosylation?
Cellular NAD+ levels, controlled by biosynthetic enzymes, directly influence the rate of ADP-ribosylation.
What CRISPR models are available for studying this modification?
Knockout, point mutation, knock-in, and overexpression models can be custom-generated.
Can EDITGENE help with my research on this topic?
Yes, EDITGENE offers a full range of CRISPR services and bioinformatics support tailored to your needs.
Conclusion
NAD+-histone H2BE35 glutamate ADP-ribosyltransferase activity (GO:0140822) is a key molecular function that bridges NAD+ metabolism and epigenetic regulation. Its dysregulation is implicated in cancer, neurodegeneration, and inflammation, making it an attractive target for therapeutic intervention. Advanced CRISPR models and multi-omics approaches are essential to fully understand its mechanisms and physiological roles. EDITGENE provides the tools and expertise to accelerate this research.
References
- 1. Zhang S et al.. 2021. Nonpeptidergic neurons suppress mast cells via glutamate to maintain skin homeostasis.. Cell 184(8):2151-2166.e16 PMID: 33765440
- 2. Zhou W et al.. 2022. SENP1-Sirt3 signaling promotes α-ketoglutarate production during M2 macrophage polarization.. Cell Rep 39(2):110660 PMID: 35417703
