SARS-CoV-2 ORF7a promotes release of inflammatory cytokines by targeting IKKβ to activate NF-κB signaling pathway
10.3969/j.issn.1000-484X.2024.04.007
- VernacularTitle:SARS-CoV-2 ORF7a通过靶向IKKβ激活NF-κB信号通路促进细胞炎症因子释放
- Author:
Lumin MOU
1
;
Qizhou LONG
;
Dongqing DENG
;
Jinzhi CHENG
;
Ying NIE
;
Jiahong WU
Author Information
1. 贵州医科大学公共卫生与健康学院环境污染监测与疾病控制重点实验室,贵阳 550025
- Keywords:
SARS-CoV-2;
ORF7a;
NF-κB signaling pathways;
Inflammatory
- From:
Chinese Journal of Immunology
2024;40(4):714-719
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the molecular mechanisms by which SARS-CoV-2 accessory protein ORF7a mediating NF-κB activation and thus inducing inflammatory cytokines production.Methods:Effects of ORF7a on NF-κB promoter activity was analyzed by luciferase reporter assay.qRT-PCR was used to analyze expressions of inflammatory cytokines.Effects of ORF7a on phos-phorylation and nuclear-translocation of p65 were determined by Western blot and immunofluorescence.Co-immunoprecipitation and immunofluorescence assay were performed to investigate potential target s of ORF7a.Results:Luciferase reporter assay showed that ORF7a activated NF-κB promoter activity in a dose-dependent manner(P<0.001),while has no effect on activation of AP-1 reporter gene.ORF7a significantly upregulates expressions of TNF-α,IL-1β and IL-8 mRNA levels(P<0.05).Western blot showed that ORF7a markedly increased phosphorylation of p65 protein(P<0.05)and p65 nuclear localization(P<0.01).The interaction between ORF7a and IKKβ protein of NF-κB signaling pathway was found by immunoprecipitation assay,and the co-localization of ORF7a and IKKβ was also confirmed by immunofluorescence assay.Conclusion:SARS-CoV-2 ORF7a targets IKKβ to promote active NF-κB pathway to the release of inflammatory cytokines.
