农业生物技术学报2024，Vol.32Issue(5)：1081-1092,12.DOI:10.3969/j.issn.1674-7968.2024.05.009
PPARγ激活和抑制状态下调控IFNβ表达的互作蛋白鉴定与分析
Identification and Analysis of Interacting Proteins Regulating IFNβ Expression in PPARγ Activated and Inhibited States
摘要
Abstract
Peroxisome proliferator activated receptor γ(PPARγ)is specifically overexpressed in alveolar macrophages and has metabolic and immunoregulatory roles.It is not clear which interacting proteins of PPARγ are involved in the regulation of typeⅠinterferon production induced by innate immunity.In this study,wild boar(Sus scrofa)lung cell line(WSL)with stable overexpression of PPARγ-3×Flag was constructed using lentiviral vector system.Taking this cell strain as a model,fluorescence quantitative PCR assay revealed that the addition of the PPARγ agonist rosiglitazone significantly reduced the level of interferon β(IFNβ)expression induced by stimulating with the dsDNA mimic Poly(dA:dT);In contrast,addition of the PPARγ inhibitor T0070907 significantly increased the level of IFNβ expression induced by Poly(dA:dT)stimulation.Further,the PPARγ-3×Flag protein complex regulating the above process was precipitated by immunomagnetic beads of anti-FLAG protein.The PPARγ-3×Flag protein complexes enriched in cytoplasmic proteins and nuclear proteins were digested into peptides and analyzed by liquid chromatography and tandem mass spectrometry(LC-MS/MS),respectively.The results showed that there were 101 cytoplasmic proteins and 95 nucleoproteins interacting specifically with PPARγ when the PPARγ agonist rosiglitazone was added.Under the condition of adding PPARγ inhibitor T0070907,there were 24 cytoplasmic proteins and 14 nuclear proteins that specifically interacted with PPARγ.GO and KEGG analysis showed that specific PPARγ interactions of cytoplasmic proteins and nuclear proteins were related to translation regulation.This study provides a new clue to further study the mechanism of regulating of IFNβexpression by PPARγ.关键词
过氧化物酶体增殖物激活受体γ(PPARγ)/干扰素β(IFNβ)/免疫沉淀-质谱联用/先天免疫/翻译调控Key words
Peroxisome proliferator activated receptor γ(PPARγ)/Interferon β(IFNβ)/Immunoprecipitation-mass spectrometry/Innate immunity/Translation regulation分类
农业科技引用本文复制引用
马卓媛,张仕强,杨玢,陈云龙,樊港,吴晓龙,王希音,华进联,王妍,张永强..PPARγ激活和抑制状态下调控IFNβ表达的互作蛋白鉴定与分析[J].农业生物技术学报,2024,32(5):1081-1092,12.基金项目
国家自然科学基金(32072856) （32072856）
国家级大学生创新创业训练计划项目(202210712123) （202210712123）
