Nuclear factor kappa B/p65 plays a positive role in peroxisome proliferator-activated receptor δ expression in orange-spotted grouper Epinephelus coioides

• Published in: Fish & shellfish immunology Vol. 102; pp. 101 - 107
• Main Authors: Yang, Min, Jinpeng, Chen, Wang, Yuxin, Wang, Qing, Wang, Shaowen, Wei, Shina, Qin, Qiwei
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.07.2020
• Subjects: Amino Acid Sequence; Animals; Bass - genetics; Bass - immunology; EcPPARδ; Epinephelus coioides; Fish Diseases - immunology; Fish Proteins - chemistry; Fish Proteins - genetics; Fish Proteins - immunology; Gene Expression Profiling - veterinary; Gene Expression Regulation - immunology; Immune responses; Immunity, Innate - genetics; Lipopolysaccharides - pharmacology; NF-kappa B - chemistry; NF-kappa B - genetics; NF-kappa B - immunology; NF-κB/p65; Nodaviridae - physiology; Poly I-C - pharmacology; PPAR delta - chemistry; PPAR delta - genetics; PPAR delta - immunology; Promoter activity; RNA Virus Infections - immunology; RNA Virus Infections - veterinary; Sequence Alignment - veterinary; Transcriptional regulation; Zymosan - pharmacology; Epinephelus coioides; NF-κB/p65; Immune responses; Transcriptional regulation; Promoter activity; EcPPARδ
• ISSN: 1050-4648; 1095-9947
• DOI: 10.1016/j.fsi.2020.03.060

The transcription factor nuclear factor kappa B (NF-κB) is a critical regulator of immune and inflammatory responses with crucial roles in various pathophysiologic conditions involving cell survival and death. Recent studies in mammals showed that NF-κB was also involved in peroxisome proliferator-activated receptors (PPARs)-mediated immune responses However, the mechanism by which NF-κB regulates PPARδ in teleosts remains unclear. In the present study, we analyzed the potential role of NF-κB/p65 (Ecp65) in the immune response stimulated by various pathogens in the grouper Epinephelus coioides. Ecp65 expression was significantly induced soon after infection with lipopolysaccharide, nervous necrosis virus, poly(I:C), and zymosan A. We also analyzed the promoter to determine the regulatory effect of Ecp65 on PPARδ expression, using progressive EcPPARδ promoter deletion mutations. Among the five truncated mutants, the luciferase reporter activity of the PPARδ-5 promoter region was highest in response to Ecp65, indicating that the core p65-binding region was located in the PPARδ-5 promoter region (+122 bp to +383 bp). Mutation analyses indicated that the luciferase reporter activity of the EcPPARδ promoter was dramatically decreased by mutation of the M3 (+305 bp to +324 bp) and M4 (+346 bp to +365 bp) binding sites, respectively. We further confirmed that Ecp65 bound to the M3 and M4 binding sites in the 5′-untranslated region of EcPPARδ by electrophoretic mobility shift assay. Finally, overexpression of Ecp65 in vitro notably promoted the transcription of EcPPARδ, interferon-related genes, and several inflammatory cytokines. This study demonstrated that Ecp65 plays an important role in modulating the innate immune responses in groupers. These results also further our understanding of the mechanisms involved in the transcriptional regulation of PPARs by p65 in bony fish. •We analyzed role of Ecp65 in the immune response against various pathogens.•Ecp65 could bind to the promoter of EcPPAR-δ and regulate its transcription.•Ecp65 could promote the transcription level of many cytokines in grouper cells.