title: P2X7 receptor positively regulates MyD88-dependent NF-κB activation.
abstract: Recent studies have demonstrated that P2X7 plays a critical role in the immune system. Here, our results showed that P2X7 activated a NF-κB - but not an IFN-β-dependent luciferase reporter gene in HEK293T cells. P2X7 was involved in the LPS- and ATP-induced NF-κB activation but did not significantly impact the response to Zymosan in RAW264.7 cells. The activation of NF-κB and IFN-β induced by myeloid differentiation primary-response protein 88 (MyD88) was enhanced by P2X7 co-expression. The siRNA silencing MyD88 almost abolished the NF-κB activation induced by P2X7, and co-immunoprecipitation showed that P2X7 interacted with MyD88. The amino acids in the C-terminus, especially the LPS-binding region of P2X7, were critical for the cellular localization and immune function of P2X7. P2X7ΔC (190 amino acids deleted in the C-terminus) and P2X7 G586A variants localized throughout the cytoplasma with a little aggregation, which differs from the cell membrane localization of wild type P2X7. Both of them could not localize to Golgi or endoplasmic reticulum. P2X7ΔC and P2X7 G586A had impaired proteolytic cleavage of caspase-1 into the functional p20 subunit, which can activate pro-inflammatory cytokines such as IL-1β. P2X7 G586A also showed a slight interaction with MyD88 in our co-immunoprecipitation experiment. This interaction might result in the attenuated activation of NF-κB and IFN-β induced by MyD88.
authors.affiliation: Key Laboratory of Regenerative Biology, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
authors.forename: Yanhui
authors.initials: Y
authors.lastname: Liu
authors.forename: Yajuan
authors.initials: Y
authors.lastname: Xiao
authors.forename: Zhiyuan
authors.initials: Z
authors.lastname: Li
mesh_terms: D000818:Animals; D020868:Gene Silencing; D017930:Genes, Reporter; D006056:Golgi Apparatus / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D016899:Interferon-beta / Q000378:metabolism; D051379:Mice; D053594:Myeloid Differentiation Factor 88 / Q000235:genetics / Q000276:immunology*; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D058486:Receptors, Purinergic P2X7 / Q000235:genetics / Q000276:immunology*; D015398:Signal Transduction / Q000276:immunology
chemical_list: D053594:Myeloid Differentiation Factor 88; D016328:NF-kappa B; D058486:Receptors, Purinergic P2X7; D016899:Interferon-beta