GO:1903073 negative regulation of death-inducing signaling complex assembly: Apoptosis Control, Genes, Functions and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:1903073 describes any process that stops, prevents, or reduces the assembly of the death-inducing signaling complex (DISC), a key platform for extrinsic apoptosis.
• DISC assembly is initiated when death ligands such as FasL or TRAIL bind their receptors, recruiting FADD and procaspase-8/10.
• Negative regulators of DISC assembly include ezrin, EMILIN2, and components of the ripoptosome, which interfere with death receptor signaling.
• Mitogen-activated protein kinase (MAPK) and retinoic acid signaling can suppress DISC formation in T cells, linking this process to immune regulation.
• Dysregulation of DISC assembly contributes to cancer, autoimmune diseases, and neurodegeneration, making it a therapeutic target.
• CRISPR-based models (knockout, knock-in, overexpression) enable functional dissection of genes that negatively regulate DISC assembly.
Description
The death-inducing signaling complex (DISC) is a multiprotein platform that assembles at the cytoplasmic tail of death receptors such as Fas (CD95) and TRAIL receptors upon ligand binding. DISC assembly is a critical step in the extrinsic apoptotic pathway, leading to caspase-8 activation and subsequent cell death. The Gene Ontology term GO:1903073, negative regulation of death-inducing signaling complex assembly, encompasses any process that stops, prevents, or reduces the frequency, rate, or extent of DISC assembly. This regulatory mechanism is essential for controlling apoptosis and preventing inappropriate cell death or survival. Researchers study this term to understand how cells fine-tune death receptor signaling in cancer, autoimmunity, and neurodegeneration.
negative regulation of death-inducing signaling complex assembly At A Glance
| GO ID | GO:1903073 |
|---|---|
| GO term | negative regulation of death-inducing signaling complex assembly |
| Ontology | biological_process |
| Synonym | negative regulation of DISC assembly; downregulation of DISC formation; inhibition of death-inducing signaling complex assembly |
| Major function | Suppression of DISC assembly to control extrinsic apoptosis |
| Related processes | Extrinsic apoptotic signaling pathway, death receptor signaling, caspase activation |
| Key regulators | Ezrin, EMILIN2, ripoptosome components, MAPK signaling |
| Disease relevance | Cancer, autoimmune diseases, neurodegeneration |
What Is GO:1903073?
GO:1903073 is a biological process term defined as any process that stops, prevents, or reduces the frequency, rate, or extent of death-inducing signaling complex assembly. In simpler terms, it covers the cellular strategies that block or dampen the formation of the DISC, a protein complex that triggers apoptosis when death receptors are activated.
Why Is negative regulation of death-inducing signaling complex assembly Important in Cell Biology?
Negative regulation of DISC assembly is crucial for maintaining cellular homeostasis and preventing aberrant apoptosis. Excessive DISC formation can lead to tissue damage in autoimmune diseases, while insufficient DISC assembly contributes to cancer cell survival. Understanding this process provides insights into therapeutic strategies for modulating cell death in various pathologies.
• Controls the threshold for extrinsic apoptosis, influencing cell fate decisions.
• Prevents inappropriate activation of caspase-8 in the absence of death ligands.
• Modulates immune responses by regulating T cell sensitivity to Fas-induced apoptosis.
• Involved in cancer resistance to TRAIL-induced apoptosis.
• EMILIN2, an extracellular matrix protein, negatively regulates DISC assembly and affects tumor progression.
• Ezrin acts as a negative regulator of death receptor-induced apoptosis.
• Dysregulation is linked to autoimmune lymphoproliferative syndromes and neurodegenerative disorders.
• Provides targets for therapeutic intervention in cancers and inflammatory diseases.
• Key for understanding the interplay between survival signaling and apoptosis.
• Enables researchers to study crosstalk between MAPK pathways and death receptor signaling.
What Happens During negative regulation of death-inducing signaling complex assembly?
Initiation of DISC assembly and its blockade
In simple terms: When death ligands bind their receptors, a complex called DISC tries to form; negative regulators step in to stop it.
DISC assembly begins with ligand-induced trimerization of death receptors such as Fas or TRAIL receptors, leading to recruitment of the adaptor protein FADD and procaspase-8/10. Negative regulation of this step can occur through proteins that interfere with receptor-ligand interactions or prevent FADD recruitment. For example, ezrin has been shown to negatively regulate death receptor-induced apoptosis by affecting DISC formation. Similarly, EMILIN2, an extracellular matrix glycoprotein, inhibits DISC assembly and subsequent caspase-8 activation.
Role of intracellular signaling pathways
In simple terms: Survival signals inside the cell can block the formation of the death complex.
Mitogen-activated protein kinase (MAPK) signaling, particularly the ERK pathway, can abrogate TRAIL-induced apoptosis upstream of caspase-8 by suppressing DISC assembly in activated T cells. Retinoic acid signaling also regulates Fas-induced apoptosis by reversing mitogen-mediated repression of DISC assembly. These pathways highlight how cellular context and external cues influence DISC formation.
Ripoptosome as a regulatory platform
In simple terms: A larger signaling platform called the ripoptosome can influence whether DISC forms or not.
The ripoptosome, a cytosolic platform that assembles in response to genotoxic stress and loss of IAPs, contains components that can modulate DISC assembly. It can sequester or modify proteins involved in DISC formation, thereby acting as a negative regulator. This crosstalk between intrinsic and extrinsic apoptotic pathways underscores the complexity of DISC regulation.
Post-translational modifications and protein interactions
In simple terms: Chemical tags and protein partnerships can turn off the death complex assembly.
Phosphorylation, ubiquitination, and other post-translational modifications of DISC components can prevent or reduce assembly. For instance, protein kinase-mediated decisions between life and death often involve phosphorylation of FADD or caspase-8, affecting DISC stability. Additionally, protein-protein interactions that mask death domains can inhibit DISC formation.
Key Genes Involved in GO:1903073 negative regulation of death-inducing signaling complex assembly
The following genes and proteins are key players in the negative regulation of death-inducing signaling complex assembly, based on published literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| FAS | Death receptor that initiates DISC assembly | Target for studying extrinsic apoptosis regulation |
| FADD | Adaptor protein recruited to death receptors | Central to DISC formation and its negative regulation |
| CASP8 | Initiator caspase activated at DISC | Effector of apoptosis; its activation is blocked by negative regulators |
| EZR | Ezrin, a negative regulator of death receptor-induced apoptosis | Modulates DISC assembly and apoptosis sensitivity |
| EMILIN2 | Extracellular matrix glycoprotein that inhibits DISC assembly | Regulates extrinsic apoptotic pathway in tumors |
| MAPK1 | ERK2, involved in MAPK signaling that suppresses DISC assembly | Links survival signaling to apoptosis resistance |
| MAPK3 | ERK1, similar to MAPK1 | Modulates TRAIL-induced apoptosis |
| RIPK1 | Component of ripoptosome, can influence DISC assembly | Crosstalk between intrinsic and extrinsic apoptosis |
| CFLAR | Caspase-8 and FADD-like apoptosis regulator (cFLIP) | Inhibits caspase-8 activation at DISC |
| TNFRSF10A | TRAIL receptor 1, initiates DISC assembly | Target for cancer therapy |
| TNFRSF10B | TRAIL receptor 2, initiates DISC assembly | Target for cancer therapy |
| BIRC2 | cIAP1, loss leads to ripoptosome formation | Regulates DISC assembly via ripoptosome |
| BIRC3 | cIAP2, similar to BIRC2 | Modulates ripoptosome and DISC |
| ATRA | Retinoic acid, regulates Fas-induced apoptosis | Reverses mitogen-mediated repression of DISC assembly |
| PTPN13 | Protein tyrosine phosphatase, potential DISC regulator | May dephosphorylate DISC components |
| SQSTM1 | p62, involved in signaling complexes | Potential role in DISC regulation |
| OPTN | Optineurin, involved in NF-kB and apoptosis | May influence DISC assembly |
How Is negative regulation of death-inducing signaling complex assembly Regulated?
The negative regulation of DISC assembly is controlled by multiple signaling pathways. MAPK/ERK signaling in activated T cells abrogates TRAIL-induced apoptosis upstream of caspase-8 by suppressing DISC formation. Retinoic acid can reverse mitogen-mediated repression of Fas DISC assembly, indicating that nuclear receptor signaling modulates this process. Additionally, the ripoptosome, which assembles upon genotoxic stress and IAP depletion, can interfere with DISC assembly through protein-protein interactions. These regulatory layers ensure that apoptosis is tightly controlled in response to environmental cues.
negative regulation of death-inducing signaling complex assembly and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| EZR | Cancer resistance to apoptosis | Ezrin knockout cancer cell lines |
| EMILIN2 | Tumor progression and apoptosis evasion | EMILIN2 overexpression in melanoma cells |
| MAPK1 | Autoimmunity and T cell apoptosis | MAPK1 knockout Jurkat T cells |
| RIPK1 | Neurodegeneration and inflammation | RIPK1 knock-in mouse models |
| CFLAR | Cancer and autoimmune lymphoproliferation | cFLIP overexpression in lymphoma cells |
Cancer
Many cancers evade apoptosis by upregulating negative regulators of DISC assembly. For example, EMILIN2 inhibits DISC assembly and its loss may contribute to tumor progression. Ezrin overexpression has been linked to resistance to death receptor-induced apoptosis in cancer cells. Understanding these mechanisms can inform targeted therapies that restore DISC formation.
Autoimmune diseases
Defective negative regulation of DISC assembly can lead to excessive apoptosis or impaired lymphocyte homeostasis, contributing to autoimmune lymphoproliferative syndromes. Retinoic acid and MAPK signaling defects may alter DISC assembly in T cells, affecting immune tolerance.
Neurodegeneration
Aberrant DISC assembly and its regulation have been implicated in neuronal cell death. Negative regulators that suppress DISC formation may protect against neurodegeneration, while their dysfunction could exacerbate apoptosis.
From negative regulation of death-inducing signaling complex assembly-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does ezrin negatively regulate DISC assembly? | EZR knockout cell lines |
| How does EMILIN2 inhibit DISC formation? | EMILIN2 overexpression and knockdown |
| What is the role of MAPK in DISC suppression? | MAPK1/3 knockout T cells |
| How does retinoic acid modulate DISC assembly? | Jurkat T cells treated with retinoic acid |
| Does ripoptosome crosstalk with DISC? | RIPK1 knockout cells with IAP depletion |
| Can point mutations in FADD affect DISC regulation? | FADD knock-in mutations |
How to Study the negative regulation of death-inducing signaling complex assembly Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Immunoprecipitation + mass spectrometry | DISC composition and interacting proteins | Identifying negative regulators |
| Caspase-8 activity assay | Caspase-8 activation downstream of DISC | Functional readout of DISC assembly |
| Annexin V flow cytometry | Apoptosis rate | Evaluating cell death |
| FRET | DISC assembly dynamics | Live-cell imaging |
| Proximity ligation assay | Protein-protein interactions in DISC | Visualizing DISC formation |
| CRISPR knockout screen | Genes affecting DISC assembly | Discovery of novel regulators |
| RNA-seq | Transcriptional changes upon DISC modulation | Pathway analysis |
| Western blot | Protein cleavage and phosphorylation | Validation of apoptosis markers |
Proteomic analysis of DISC
Immunoprecipitation of death receptors followed by mass spectrometry can identify DISC components and their negative regulators. This method reveals dynamic changes in complex composition upon stimulation.
Apoptosis assays
Flow cytometry with Annexin V/propidium iodide staining and caspase-8 activity assays measure the functional outcome of DISC regulation. These assays are used to assess how genetic perturbations affect apoptosis.
Imaging of DISC assembly
Fluorescence resonance energy transfer (FRET) and proximity ligation assays visualize DISC formation at the single-cell level. These techniques help localize negative regulators in real time.
CRISPR screening
Genome-wide CRISPR knockout screens can identify genes whose loss enhances or suppresses DISC assembly. This approach uncovers novel negative regulators.
How CRISPR Can Be Used to Study GO:1903073 negative regulation of death-inducing signaling complex assembly
Knockout
CRISPR knockout of candidate negative regulators such as EZR or EMILIN2 can test their role in DISC assembly. Loss of function may enhance DISC formation and sensitize cells to apoptosis.
Point Mutation
Introducing point mutations in genes like FADD or CASP8 can dissect specific residues required for negative regulation. This approach reveals phosphorylation or interaction sites critical for DISC control.
Knock-in
Knock-in of tagged versions of DISC components (e.g., GFP-FADD) allows real-time tracking of complex assembly. This helps visualize how negative regulators affect DISC dynamics.
Overexpression
Overexpression of negative regulators like EMILIN2 or ezrin can suppress DISC assembly and confer apoptosis resistance. This models cancer-associated overexpression.
How EDITGENE Supports negative regulation of death-inducing signaling complex assembly Research
Researchers studying negative regulation of death-inducing signaling complex assembly-related genes often need to determine whether a candidate gene is causally involved in modulating DISC formation. EDITGENE provides comprehensive CRISPR-based services to accelerate this research.
Contact EDITGENE today to design your custom CRISPR model for negative regulation of death-inducing signaling complex assembly research.
Frequently Asked Questions About negative regulation of death-inducing signaling complex assembly
What is GO:1903073?
GO:1903073 is a Gene Ontology term for any process that stops, prevents, or reduces the frequency, rate, or extent of death-inducing signaling complex (DISC) assembly.
What genes are involved in negative regulation of DISC assembly?
Key genes include EZR, EMILIN2, MAPK1, MAPK3, RIPK1, and CFLAR, among others.
How is DISC assembly negatively regulated?
Through proteins like ezrin and EMILIN2, signaling pathways like MAPK/ERK, and platforms like the ripoptosome that interfere with DISC formation.
Why is negative regulation of DISC assembly important in cancer?
It allows cancer cells to evade apoptosis; upregulation of negative regulators like EMILIN2 or ezrin contributes to resistance to death receptor-induced cell death.
What experimental models are used to study DISC assembly?
Knockout cell lines, overexpression models, point mutations, and CRISPR screens are commonly used.
Can retinoic acid affect DISC assembly?
Yes, retinoic acid regulates Fas-induced apoptosis in Jurkat T cells by reversing mitogen-mediated repression of DISC assembly.
What is the role of MAPK in DISC regulation?
MAPK/ERK signaling in activated T cells abrogates TRAIL-induced apoptosis upstream of caspase-8 by suppressing DISC assembly.
How does the ripoptosome relate to DISC?
The ripoptosome can modulate DISC assembly through protein interactions, especially under genotoxic stress and IAP depletion.
What diseases are linked to dysregulated DISC assembly?
Cancer, autoimmune diseases, and neurodegeneration are associated with altered DISC regulation.
How can CRISPR help study negative regulation of DISC assembly?
CRISPR knockout, knock-in, point mutation, and overexpression models enable functional dissection of genes controlling DISC assembly.
Conclusion
GO:1903073, negative regulation of death-inducing signaling complex assembly, is a critical process that fine-tunes extrinsic apoptosis. Its dysregulation contributes to cancer, autoimmunity, and neurodegeneration. Understanding the molecular players and pathways involved offers therapeutic opportunities. EDITGENE provides advanced CRISPR tools to accelerate research in this field.
References
- 1. Engin A. 2021. Protein Kinase-Mediated Decision Between the Life and Death.. Adv Exp Med Biol 1275:1-33 PMID: 33539010
- 2. Kuo WC et al.. 2010. Ezrin is a negative regulator of death receptor-induced apoptosis.. Oncogene 29(9):1374-83 PMID: 19935704
- 3. Mongiat M et al.. 2007. Regulation of the extrinsic apoptotic pathway by the extracellular matrix glycoprotein EMILIN2.. Mol Cell Biol 27(20):7176-87 PMID: 17698584
- 4. Engedal N et al.. 2009. Retinoic acid regulates Fas-induced apoptosis in Jurkat T cells: reversal of mitogen-mediated repression of Fas DISC assembly.. J Leukoc Biol 85(3):469-80 PMID: 19112091
- 5. Tenev T et al.. 2011. The Ripoptosome, a signaling platform that assembles in response to genotoxic stress and loss of IAPs.. Mol Cell 43(3):432-48 PMID: 21737329
- 6. Söderström TS et al.. 2002. Mitogen-activated protein kinase/extracellular signal-regulated kinase signaling in activated T cells abrogates TRAIL-induced apoptosis upstream of the mitochondrial amplification loop and caspase-8.. J Immunol 169(6):2851-60 PMID: 12218097
