Related Experiment Video
Updated: Jun 1, 2026
10:57
NF-κB-dependent Luciferase Activation and Quantification of Gene Expression in Salmonella Infected Tissue Culture Cells
Published on: January 12, 2020
P2X7 receptor positively regulates MyD88-dependent NF-κB activation
Yanhui Liu1, Yajuan Xiao, Zhiyuan Li
1Key Laboratory of Regenerative Biology, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
Cytokine
|May 31, 2011
Summary
The P2X7 receptor activates NF-κB signaling, interacting with MyD88 to regulate immune responses. Its C-terminus is crucial for localization and function, impacting caspase-1 cleavage and cytokine release.
Area of Science:
- Immunology
- Cell Biology
- Molecular Biology
Background:
- P2X7 receptor is recognized for its significant role in immune system regulation.
- Understanding P2X7's molecular mechanisms is key to deciphering immune signaling pathways.
Purpose of the Study:
- To elucidate the role of P2X7 in NF-κB and IFN-β activation.
- To investigate the interaction between P2X7 and MyD88.
- To determine the functional significance of P2X7's C-terminus in immune responses.
Main Methods:
- Luciferase reporter gene assays in HEK293T cells.
- Stimulation assays using LPS, ATP, and Zymosan in RAW264.7 cells.
- Co-immunoprecipitation and siRNA silencing to study protein interactions and functional dependencies.
- Analysis of P2X7 variants (P2X7ΔC, P2X7 G586A) for cellular localization and caspase-1 cleavage.
Main Results:
- P2X7 activated NF-κB but not IFN-β reporter genes.
- P2X7 mediated LPS- and ATP-induced NF-κB activation, but not Zymosan-induced activation.
- P2X7 co-expression enhanced MyD88-induced NF-κB and IFN-β activation.
- P2X7 directly interacted with MyD88, and MyD88 silencing abolished P2X7-induced NF-κB activation.
- The C-terminus of P2X7, including the LPS-binding region, is critical for its localization and immune function.
- P2X7 variants lacking the C-terminus or with G586A mutation showed altered cellular localization and impaired caspase-1 cleavage.
- These variants exhibited reduced ability to activate pro-inflammatory cytokines like IL-1β.
Conclusions:
- P2X7 signaling is dependent on MyD88 for NF-κB activation.
- The C-terminus of P2X7 is essential for its proper localization, interaction with MyD88, and downstream immune functions, including caspase-1 activation and cytokine production.
- These findings highlight P2X7's intricate role in innate immunity and inflammatory signaling.
Related Concept Videos
02:26NF-κB-dependent Signaling Pathway
The transcription factor NF-κB was discovered in 1986 in the lab of Nobel laureate Professor David Baltimore, for its interaction with the immunoglobulin light chain enhancer in B-cells. After more than three decades of study, it is now evident that NF-κB regulates the expression of over 100 genes. Most of these genes play an essential role in the innate and adaptive immune responses as well as the inflammatory responses of animals.
NF-κB-dependent Signaling Mechanism
The heterodimer of NF-κB...
NF-κB-dependent Signaling Mechanism
The heterodimer of NF-κB...
02:26NF-kB-dependent Signaling Pathway
The transcription factor NF-κB was discovered in 1986 in the lab of Nobel laureate Professor David Baltimore, for its interaction with the immunoglobulin light chain enhancer in B-cells. After more than three decades of study, it is now evident that NF-κB regulates the expression of over 100 genes. Most of these genes play an essential role in the innate and adaptive immune responses as well as the inflammatory responses of animals.
NF-κB-dependent Signaling Mechanism
The heterodimer of NF-κB...
NF-κB-dependent Signaling Mechanism
The heterodimer of NF-κB...
02:04Co-activators and Co-repressors
Gene transcription is regulated by the synergistic action of several proteins that form a complex at a gene regulatory site. This is observed in eukaryotes, where the regulation of gene expression is a complex process. Regulatory proteins in eukaryotes can broadly be classified into two types – regulators that bind directly to specific DNA sequences and co-regulators that associate with regulatory proteins but cannot directly bind to the DNA. These co-regulators are further divided into...
01:20The JAK-STAT Signaling Pathway
Several cytokine receptors have tightly bound Janus kinase or JAK proteins attached at their cytosolic tail. Small signaling molecules such as cytokines, growth hormones, or prolactins bind to the cytokine receptors and initiate their dimerization. The dimerization brings the cytosolic JAKs together that trans-phosphorylate and activates each other. The activated JAKs now phosphorylate cytosolic tails of the cytokine receptors, which serve as binding sites for adaptor proteins such as SH2...
01:07MAPK Signaling Cascades
Mitogen-activated protein kinase, or MAPK pathway, activates three sequential kinases to regulate cellular responses such as proliferation, differentiation, survival, and apoptosis. The canonical MAPK pathway starts with a mitogen or growth factor binding to an RTK. The activated RTKs stimulate Ras, which recruits Raf or MAP3 Kinase (MAPKKK), the first kinase of the MAPK signaling cascade. Raf further phosphorylates and activates MEK or MAP2 Kinases (MAPKK), which in turn phosphorylates MAP...
01:24T Cell Types and Functions
When T cells with CD4 markers are activated, they give rise to two types of effector cells: helper T cells and regulatory T cells. Meanwhile, T cells with CD8 markers differentiate into effector cytotoxic T cells. The differentiation of CD4 T cells into helper T cell subsets, such as Th1, Th2, and Th17 cells, is dependent on the antigen type, antigen-presenting cell, and regulatory cytokines.
Th1 cells stimulate dendritic cells to express necessary co-stimulatory molecules on their surfaces for...
Th1 cells stimulate dendritic cells to express necessary co-stimulatory molecules on their surfaces for...