GO:0035872 nucleotide-binding domain, leucine rich repeat containing receptor signaling pathway: Innate Immune Signaling, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0035872 describes the signaling pathway mediated by nucleotide-binding domain, leucine rich repeat containing receptors (NLRs), which are cytosolic sensors that assemble inflammasomes and initiate innate immune responses.
• NLR signaling is central to host defense, inflammation, and cell death, and its dysregulation is linked to cancer, autoinflammatory diseases, and neurological disorders.
• Key NLR family members include NLRP1, NLRP3, NLRC4, and NAIP, which form multiprotein inflammasome complexes upon ligand sensing.
• The pathway involves ligand recognition, inflammasome assembly, caspase-1 activation, cytokine maturation (IL-1β, IL-18), and pyroptotic cell death.
• NLR signaling crosstalks with autophagy, mitochondrial function, and other innate immune pathways, influencing disease outcomes.
• CRISPR-based models (knockout, knock-in, point mutation) are essential for dissecting NLR gene function and for developing targeted therapeutics.
Description
The nucleotide-binding domain, leucine rich repeat containing receptor (NLR) signaling pathway (GO:0035872) is a biological process that governs cytosolic innate immune sensing and downstream inflammatory responses. NLRs are a family of intracellular pattern recognition receptors that detect pathogen-associated molecular patterns (PAMPs) and danger-associated molecular patterns (DAMPs), leading to the assembly of inflammasomes and activation of inflammatory caspases. This pathway is critical for host defense against microbial infections and for maintaining tissue homeostasis. Dysregulated NLR signaling contributes to a wide range of human diseases, including cancer, autoinflammatory disorders, and neurodegeneration. Understanding the molecular mechanisms of NLR signaling is therefore essential for developing targeted therapies and for interpreting genetic variants in immune-related genes.
nucleotide-binding domain, leucine rich repeat containing receptor signaling pathway At A Glance
| GO ID | GO:0035872 |
|---|---|
| GO term | nucleotide-binding domain, leucine rich repeat containing receptor signaling pathway |
| Ontology | biological_process |
| Synonym | None listed in QuickGO |
| Major function | Cytosolic sensing of PAMPs/DAMPs and initiation of inflammatory and cell death responses via inflammasome assembly |
| Key receptors | NLRP1, NLRP3, NLRC4, NAIP, and other NLR family members |
| Downstream effectors | Caspase-1, IL-1β, IL-18, gasdermin D |
| Crosstalk | Autophagy, mitochondrial signaling, and other innate immune pathways |
| Disease relevance | Cancer, autoinflammatory diseases, neurodegeneration, metabolic disorders |
What Is GO:0035872?
GO:0035872 defines the series of molecular events initiated by nucleotide-binding domain, leucine rich repeat containing receptors (NLRs) that transmit signals from cytosolic ligands to downstream effectors, typically resulting in inflammasome assembly, caspase-1 activation, and the processing of pro-inflammatory cytokines such as IL-1β and IL-18. This pathway is a key component of innate immunity and is conserved across vertebrates.
Why Is nucleotide-binding domain, leucine rich repeat containing receptor signaling pathway Important in Cell Biology?
The NLR signaling pathway is a cornerstone of innate immunity, providing rapid detection of microbial invaders and cellular stress. Its proper regulation is essential for clearing infections and for preventing excessive inflammation that can damage host tissues. Moreover, NLR signaling influences adaptive immunity, tumor surveillance, and neurological function, making it a high-priority target for therapeutic intervention.
• Mediates host defense against bacterial, viral, and fungal pathogens through inflammasome activation.
• Drives maturation and secretion of pro-inflammatory cytokines IL-1β and IL-18, which are central to fever and inflammation.
• Induces pyroptosis, a lytic form of cell death that restricts pathogen replication and releases alarmins.
• Plays a dual role in cancer: promoting anti-tumor immunity or fostering tumor-promoting inflammation depending on context.
• Implicated in autoinflammatory diseases such as cryopyrin-associated periodic syndromes (CAPS) caused by NLRP3 mutations.
• Contributes to neuroinflammation and neuronal pyroptosis after intracerebral hemorrhage via CCR5/PKA/CREB signaling.
• Crosstalks with autophagy, which controls intestinal epithelial barrier integrity and inflammation.
• Mitochondrial function and reactive oxygen species modulate NLRP3 inflammasome activation.
• Genetic variants in NLR genes are associated with susceptibility to inflammatory and autoimmune disorders.
• CRISPR screening identifies novel regulators of NLR signaling, offering new drug targets.
What Happens During nucleotide-binding domain, leucine rich repeat containing receptor signaling pathway?
Ligand Recognition and Receptor Activation
In simple terms: NLR proteins act like security guards inside the cell, detecting signs of infection or danger.
NLRs contain a central nucleotide-binding domain (NBD) and C-terminal leucine-rich repeats (LRRs) that autoinhibit the receptor in the absence of ligand. Upon binding to specific PAMPs or DAMPs, the LRR domain undergoes conformational changes that relieve autoinhibition, allowing oligomerization and activation. For example, NLRP3 senses diverse stimuli including ATP, nigericin, and crystalline substances, while NLRC4 detects bacterial flagellin and type III secretion system components.
Inflammasome Assembly and Caspase-1 Activation
In simple terms: Activated NLRs team up with an adaptor protein to form a molecular platform that turns on an inflammatory enzyme.
Activated NLRs recruit the adaptor protein ASC (PYCARD) via homotypic pyrin domain interactions, leading to ASC speck formation and recruitment of pro-caspase-1. This multiprotein complex, termed the inflammasome, facilitates proximity-induced autoproteolysis of caspase-1, generating active caspase-1. Active caspase-1 then cleaves pro-IL-1β and pro-IL-18 into their mature forms and cleaves gasdermin D to trigger pyroptosis.
Cytokine Maturation and Pyroptosis
In simple terms: The active enzyme cuts inactive cytokine precursors into active messengers and punches holes in the cell membrane, causing a controlled burst of inflammation.
Mature IL-1β and IL-18 are secreted through gasdermin D pores, amplifying inflammation and recruiting immune cells. Gasdermin D cleavage also induces pyroptosis, a lytic cell death that releases additional DAMPs and cytokines, further propagating the immune response. This process is critical for clearing intracellular pathogens but can cause tissue damage if unchecked.
Crosstalk with Autophagy and Mitochondrial Signaling
In simple terms: The NLR pathway talks to other cellular cleanup and energy systems to keep inflammation in check.
Autophagy negatively regulates inflammasome activation by targeting NLRs and inflammasome components for degradation, thereby limiting excessive inflammation. Mitochondria contribute to NLRP3 activation by producing reactive oxygen species and releasing mitochondrial DNA, which serve as danger signals. This crosstalk is essential for maintaining intestinal epithelial barrier function and preventing chronic inflammatory diseases.
Key Genes Involved in GO:0035872 nucleotide-binding domain, leucine rich repeat containing receptor signaling pathway
The following genes encode key components of the NLR signaling pathway, including receptors, adaptors, and effectors, which are frequently studied using CRISPR-based models.
| Gene | Major Role | Research Relevance |
|---|---|---|
| NLRP1 | Forms inflammasome in response to bacterial toxins and danger signals; activates caspase-1 | Studied in neuronal pyroptosis after intracerebral hemorrhage and cancer |
| NLRP3 | Senses diverse PAMPs/DAMPs; assembles inflammasome with ASC and caspase-1 | Target for autoinflammatory diseases and metabolic disorders |
| NLRC4 | Detects bacterial flagellin and T3SS components; forms inflammasome with NAIP | Key for host defense against Salmonella and Legionella |
| NAIP | Co-receptor for NLRC4; binds bacterial ligands directly | Determines ligand specificity of NLRC4 inflammasome |
| ASC (PYCARD) | Adaptor linking NLRs to caspase-1; forms specks | Central node for inflammasome assembly; knockout blocks cytokine maturation |
| CASP1 | Effector caspase; cleaves IL-1β, IL-18, and gasdermin D | Essential for pyroptosis and cytokine release |
| IL1B | Pro-inflammatory cytokine matured by caspase-1 | Biomarker and therapeutic target in inflammatory diseases |
| IL18 | Pro-inflammatory cytokine matured by caspase-1 | Implicated in autoinflammatory and autoimmune conditions |
| GSDMD | Gasdermin D; forms pores upon cleavage by caspase-1 | Executes pyroptosis; knockout prevents cell lysis |
| CCR5 | Chemokine receptor; activates NLRP1-dependent pyroptosis via PKA/CREB | Potential target for neuroinflammation after stroke |
| PKA | Protein kinase A; modulates NLRP1 activation downstream of CCR5 | Signaling node in neuronal pyroptosis |
| CREB | Transcription factor activated by PKA; regulates NLRP1 expression | Links chemokine signaling to inflammasome activation |
| ATG5 | Autophagy-related gene; regulates inflammasome degradation | Modulates intestinal inflammation and barrier function |
| ATG7 | Autophagy-related gene; controls autophagosome formation | Impacts NLRP3 stability and inflammation |
| MTOR | Kinase that inhibits autophagy; indirectly promotes inflammasome activity | Target for modulating NLR signaling in disease |
| TXNIP | Thioredoxin-interacting protein; links oxidative stress to NLRP3 activation | Mediates mitochondrial ROS-driven inflammasome |
| VDAC | Voltage-dependent anion channel; regulates mitochondrial ROS and NLRP3 | Mitochondrial checkpoint for inflammasome |
| NLRP6 | Regulates intestinal homeostasis and antimicrobial peptide secretion | Protects against colitis and enteric infections |
How Is nucleotide-binding domain, leucine rich repeat containing receptor signaling pathway Regulated?
NLR signaling is tightly regulated at multiple levels. Autophagy acts as a negative regulator by degrading inflammasome components, and its inhibition enhances NLRP3-dependent inflammation. Mitochondrial reactive oxygen species (ROS) and mitochondrial DNA release positively regulate NLRP3 activation, while antioxidants suppress it. Post-translational modifications such as phosphorylation, ubiquitination, and SUMOylation modulate NLR activity. Additionally, the CCR5/PKA/CREB axis promotes NLRP1-dependent neuronal pyroptosis after intracerebral hemorrhage, illustrating G-protein-coupled receptor crosstalk. These regulatory mechanisms are critical for preventing excessive inflammation and are often dysregulated in disease.
nucleotide-binding domain, leucine rich repeat containing receptor signaling pathway and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| NLRP3 | Cryopyrin-associated periodic syndromes (CAPS); obesity-induced inflammation | Knock-in mouse models with CAPS-associated mutations; adipocyte-specific KO |
| NLRP1 | Neuronal pyroptosis after intracerebral hemorrhage | CCR5 KO and NLRP1 KO mice; neuronal cell lines |
| NLRC4 | Salmonella and Legionella infection susceptibility | NLRC4 KO mice; macrophage infection models |
| ASC (PYCARD) | Defective inflammasome assembly; susceptibility to infections | ASC KO mice; CRISPR KO in THP-1 cells |
| CASP1 | Impaired cytokine maturation and pyroptosis | Casp1 KO mice; human monocyte-derived macrophages |
Cancer
NLR signaling plays a dual role in cancer. Inflammasome activation can promote anti-tumor immunity by enhancing antigen presentation and cytotoxic T cell responses, but chronic inflammation driven by NLRs can also foster tumorigenesis, angiogenesis, and metastasis. For example, NLRP3 inflammasome activity in myeloid cells can suppress tumor growth in colitis-associated cancer models, while in other contexts it promotes tumor progression. Targeting NLR signaling is therefore a promising but context-dependent therapeutic strategy.
Neuroinflammation and Stroke
After intracerebral hemorrhage, CCR5 activation promotes NLRP1-dependent neuronal pyroptosis via the CCR5/PKA/CREB pathway, contributing to secondary brain injury. Inhibition of this axis reduces neuronal death and improves outcomes in preclinical models, suggesting that NLR signaling is a viable target for neuroprotective therapies. Additionally, NLRP3 inflammasome activation is implicated in Alzheimer's disease and multiple sclerosis, where it drives chronic neuroinflammation.
Autoinflammatory and Autoimmune Diseases
Gain-of-function mutations in NLRP3 cause cryopyrin-associated periodic syndromes (CAPS), characterized by recurrent fever and inflammation. Similarly, dysregulated NLR signaling is associated with inflammatory bowel diseases, where autophagy defects exacerbate inflammasome activation and compromise the intestinal epithelial barrier. Plant autoimmunity studies have also revealed conserved NLR signaling principles, underscoring the evolutionary importance of this pathway.
Metabolic Disorders
Obesity-induced adipose tissue inflammation involves NLRP3 inflammasome activation, which contributes to insulin resistance and type 2 diabetes. Mitochondrial dysfunction and ROS production in adipocytes activate NLRP3, linking metabolic stress to chronic inflammation. Modulating NLR signaling may therefore have therapeutic potential in metabolic diseases.
From nucleotide-binding domain, leucine rich repeat containing receptor signaling pathway-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does NLRP3 drive obesity-induced insulin resistance? | Adipose-specific NLRP3 knockout mouse |
| What is the role of CCR5 in neuronal pyroptosis? | CCR5 knockout and NLRP1 knockout mice |
| How does autophagy regulate inflammasome activity? | ATG5 or ATG7 knockout intestinal epithelial cells |
| Can a point mutation in NLRP3 mimic CAPS? | NLRP3 knock-in mice with disease-associated mutation |
| What is the impact of NLRC4 ligand recognition? | NLRC4 knockout macrophages reconstituted with mutant NLRC4 |
| Does mitochondrial ROS activate NLRP3? | TXNIP or VDAC knockout cells treated with ROS inducers |
How to Study the nucleotide-binding domain, leucine rich repeat containing receptor signaling pathway Process
| Method | What It Measures | Typical Application |
|---|---|---|
| CRISPR knockout screening | Loss-of-function effects on NLR signaling | Identify novel regulators of inflammasome activation |
| RNA-seq | Transcriptional changes in NLR and cytokine genes | Profile inflammatory responses in disease models |
| Immunoprecipitation-mass spectrometry | Protein-protein interactions of NLRs | Map inflammasome interactome |
| ASC speck imaging | Inflammasome assembly | Visualize activation in live cells |
| IL-1β ELISA | Cytokine secretion | Quantify inflammasome output |
| LDH release assay | Pyroptosis | Measure cell death |
| Western blot | Caspase-1 cleavage and gasdermin D processing | Confirm inflammasome activation |
| Flow cytometry | Immune cell activation and cytokine production | Analyze immune responses in vivo |
CRISPR Screening for NLR Pathway Regulators
Genome-wide CRISPR knockout screens can identify novel genes that regulate NLR signaling, such as those affecting inflammasome assembly or cytokine secretion. These screens typically use reporter cell lines expressing fluorescently tagged IL-1β or caspase-1 activity sensors, followed by flow cytometry or sequencing to identify enriched sgRNAs.
Transcriptomics and Proteomics
RNA-seq reveals transcriptional changes in NLR genes and downstream inflammatory mediators upon stimulation. Proteomics, including immunoprecipitation-mass spectrometry, can map interactomes of NLRs and identify post-translational modifications critical for signaling.
Imaging and Functional Assays
Live-cell imaging of ASC speck formation and gasdermin D pore formation provides spatial and temporal insights into inflammasome activation. Functional assays such as ELISA for IL-1β and LDH release for pyroptosis are standard readouts.
Animal Models and In Vivo Studies
Knockout and knock-in mouse models are essential for studying NLR signaling in physiology and disease, including infection, cancer, and neuroinflammation. These models allow assessment of survival, tissue pathology, and immune cell infiltration.
How CRISPR Can Be Used to Study GO:0035872 nucleotide-binding domain, leucine rich repeat containing receptor signaling pathway
Knockout
CRISPR knockout of NLR genes (e.g., NLRP3, NLRC4, CASP1) in cell lines and mice is widely used to dissect their roles in inflammasome activation and disease. For example, NLRP3 knockout macrophages fail to secrete IL-1β in response to ATP, confirming its essential function. Knockout models also help identify redundant or specific roles of NLR family members.
Point Mutation
Introducing disease-associated point mutations (e.g., NLRP3 R258W, associated with CAPS) via CRISPR base editing or HDR allows study of gain-of-function effects on inflammasome activity. Such models are valuable for testing targeted therapies and understanding mutation-specific signaling.
Knock-in
Knock-in of reporter tags (e.g., fluorescent proteins) or epitope tags into endogenous NLR loci enables real-time tracking of protein expression and localization. Knock-in of human NLR genes into mouse models can humanize the pathway for drug testing.
Overexpression
Overexpression of NLRs or inflammasome components using CRISPR activation (CRISPRa) or lentiviral vectors can amplify signaling for biochemical studies or screening. This approach is useful for identifying downstream effectors and for producing large quantities of proteins for structural studies.
How EDITGENE Supports nucleotide-binding domain, leucine rich repeat containing receptor signaling pathway Research
Researchers studying nucleotide-binding domain, leucine rich repeat containing receptor signaling pathway-related genes often need to determine whether a candidate gene is causally involved in inflammasome activation, cytokine production, or disease progression. EDITGENE provides comprehensive CRISPR-based services to generate precisely engineered cell and animal models, enabling rigorous functional validation of NLR pathway components.
Contact EDITGENE today to design your custom CRISPR model for nucleotide-binding domain, leucine rich repeat containing receptor signaling pathway research.
Related Products
| Product name | Cat.No. | Species | Gene ID | |
|---|---|---|---|---|
| MAP3K7 Knockout HEK293 Cell Line | EDJ-KQ142 | Human | 6885 | Details Get a Quote |
| CYLD Knockout HEK293 Cell Line | EDJ-KQ560 | Human | 1540 | Details Get a Quote |
| TNFAIP3 Knockout HEK293 Cell Line | EDJ-KQ595 | Human | 7128 | Details Get a Quote |
| MAP2K6 Knockout HEK293 Cell Line | EDJ-KQ1353 | Human | 5608 | Details Get a Quote |
| ITCH Knockout HEK293 Cell Line | EDJ-KQ1468 | Human | 83737 | Details Get a Quote |
| RNF34 Knockout HEK293 Cell Line | EDJ-KQ9480 | Human | 80196 | Details Get a Quote |
| TNFAIP3 Knockout HeLa Cell Line | EDJ-KQ18184 | Human | 7128 | Details Get a Quote |
| CYLD Knockout A-549 Cell Line | EDJ-KQ18939 | Human | 1540 | Details Get a Quote |
| CYLD Knockout HCT 116 Cell Line | EDJ-KQ18940 | Human | 1540 | Details Get a Quote |
| CYLD Knockout HeLa Cell Line | EDJ-KQ18941 | Human | 1540 | Details Get a Quote |
| MAP3K7 Knockout A-549 Cell Line | EDJ-KQ18978 | Human | 6885 | Details Get a Quote |
| MAP3K7 Knockout HCT 116 Cell Line | EDJ-KQ18980 | Human | 6885 | Details Get a Quote |
| MAP3K7 Knockout HeLa Cell Line | EDJ-KQ18981 | Human | 6885 | Details Get a Quote |
| TNFAIP3 Knockout A-549 Cell Line | EDJ-KQ19035 | Human | 7128 | Details Get a Quote |
| TNFAIP3 Knockout HCT 116 Cell Line | EDJ-KQ19036 | Human | 7128 | Details Get a Quote |
Frequently Asked Questions About nucleotide-binding domain, leucine rich repeat containing receptor signaling pathway
What is GO:0035872?
GO:0035872 is the Gene Ontology term for the nucleotide-binding domain, leucine rich repeat containing receptor signaling pathway, a biological process that mediates cytosolic innate immune sensing and inflammasome activation.
What genes are involved in nucleotide-binding domain, leucine rich repeat containing receptor signaling pathway?
Key genes include NLRP1, NLRP3, NLRC4, NAIP, ASC (PYCARD), CASP1, IL1B, IL18, and GSDMD, among others.
How does the NLR signaling pathway work?
NLRs detect PAMPs/DAMPs, undergo conformational changes, assemble inflammasomes with ASC and caspase-1, leading to cytokine maturation and pyroptosis.
What diseases are associated with NLR signaling?
Diseases include cryopyrin-associated periodic syndromes, inflammatory bowel disease, cancer, neuroinflammation, and metabolic disorders.
What is the role of NLRP3 in inflammation?
NLRP3 senses diverse danger signals and forms an inflammasome that activates caspase-1, leading to IL-1β and IL-18 release and pyroptosis.
How is NLR signaling regulated?
It is regulated by autophagy, mitochondrial ROS, post-translational modifications, and crosstalk with other signaling pathways such as CCR5/PKA/CREB.
Can CRISPR be used to study NLR signaling?
Yes, CRISPR knockout, knock-in, and point mutation models are widely used to dissect NLR gene function and disease mechanisms.
What are the main methods to study NLR signaling?
Common methods include CRISPR screening, RNA-seq, proteomics, imaging of ASC specks, cytokine ELISA, and LDH release assays.
What is the connection between autophagy and NLR signaling?
Autophagy negatively regulates inflammasomes by degrading NLRs and inflammasome components, thereby limiting inflammation.
How does mitochondrial function affect NLR signaling?
Mitochondria produce ROS and release DNA that activate NLRP3, linking metabolic stress to inflammasome activation.
Conclusion
The nucleotide-binding domain, leucine rich repeat containing receptor signaling pathway (GO:0035872) is a fundamental innate immune mechanism with broad implications for health and disease. Its dysregulation contributes to cancer, autoinflammatory disorders, neurodegeneration, and metabolic diseases, making it a prime target for therapeutic intervention. Advances in CRISPR technology and functional genomics are accelerating our understanding of this pathway and enabling the development of precision medicines.
References
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- 2. Foerster EG et al.. 2022. How autophagy controls the intestinal epithelial barrier.. Autophagy 18(1):86-103 PMID: 33906557
- 3. Engin A. 2024. Reappraisal of Adipose Tissue Inflammation in Obesity.. Adv Exp Med Biol 1460:297-327 PMID: 39287856
- 4. Yan J et al.. 2021. CCR5 Activation Promotes NLRP1-Dependent Neuronal Pyroptosis via CCR5/PKA/CREB Pathway After Intracerebral Hemorrhage.. Stroke 52(12):4021-4032 PMID: 34719258
- 5. Karki R et al.. 2017. Inflammasomes and Cancer.. Cancer Immunol Res 5(2):94-99 PMID: 28093447
- 6. Chakraborty J et al.. 2018. Autoimmunity in plants.. Planta 248(4):751-767 PMID: 30046903
- 7. Lee S et al.. 2016. Regulation and Function of the Nucleotide Binding Domain Leucine-Rich Repeat-Containing Receptor, Pyrin Domain-Containing-3 Inflammasome in Lung Disease.. Am J Respir Cell Mol Biol 54(2):151-60 PMID: 26418144
- 8. Banoth B et al.. 2018. Mitochondria in innate immune signaling.. Transl Res 202:52-68 PMID: 30165038
