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Similar to mammals, fish possess interferon (IFN) regulatory factor 2 (IRF2)-dependent type I IFN responses. Nevertheless, the detailed mechanism through which IRF2 regulates type I IFNa3 remains largely unknown. In the present study, we first identified two genes from golden pompano (Trachinotus ovatus), IRF2 (ToIRF2) and IFNa3 (ToIFNa3), in the IFN/IRF-based signalling pathway. The open reading frame (ORF) sequence of ToIRF2 encoded 335 amino acids possessing four typical characteristic domains, including a conserved DNA-binding domain (DBD), an interferon association domain 2 (IAD2), a transcriptional activation domain (TAD), and a transcriptional repression domain (TRD). Furthermore, transcripts of ToIRF2 were significantly upregulated after stimulation by polyinosinic: polycytidylic acid [poly (I:C)], lipopolysaccharide (LPS) and flagellin in immune-related tissues (blood, liver, and head-kidney). Moreover, to investigate whether ToIRF2 was a regulator of ToIFNa3, promoter analysis was performed. The results showed that the region from -896 bp to -200 bp is defined as the core promoter using progressive deletion mutations of IFNa3. Additionally, ToIRF2 overexpression led to a clear time-dependent enhancement of ToIFNa3 promoter expression in HEK293T cells. Mutation analyses indicated that the activity of the ToIFNa3 promoter significantly decreased after targeted mutation of M4/5 binding sites. Electrophoretic mobile shift assays (EMSAs) verified that IRF2 interacted with the binding site of the ToIFNa3 promoter region to regulate ToIFNa3 transcription. Last, the promoter activity of ToIFNa3-2 was more responsive to treatment with poly (I:C) than LPS and flagellin. Furthermore, overexpression of ToIRF2 in vitro obviously increased the expression of several IFN/IRF-based signalling pathway genes after poly (I:C) abduction. In conclusion, the present study provides the first evidence of the positive regulation of ToIFNa3 transcription by ToIRF2 and contributes to a better understanding of the transcriptional mechanisms of ToIRF2 in fish.

Titel:	Functional characterization of interferon regulatory factor 2 and its role in the transcription of interferon a3 in golden pompano Trachinotus ovatus (Linnaeus 1758).
Autor/in / Beteiligte Person:	Zhu, KC ; Guo, HY ; Zhang, N ; Guo, L ; Liu, BS ; Jiang, SG ; Zhang, DC
Zeitschrift:	Fish & shellfish immunology, Jg. 93 (2019-10-01), S. 90-98
Veröffentlichung:	London ; New York : Academic Press, c1991-, 2019
Medientyp:	academicJournal
ISSN:	1095-9947 (electronic)
DOI:	10.1016/j.fsi.2019.07.045
Schlagwort:	Amino Acid Sequence Animals Base Sequence Female Fish Proteins chemistry Fish Proteins genetics Fish Proteins immunology Flagellin pharmacology Gene Expression Profiling veterinary Interferon Regulatory Factor-2 chemistry Lipopolysaccharides pharmacology Male Phylogeny Poly I-C pharmacology Promoter Regions, Genetic Fish Diseases immunology Fishes genetics Fishes immunology Gene Expression Regulation immunology Immunity, Innate genetics Interferon Regulatory Factor-2 genetics Interferon Regulatory Factor-2 immunology
Sonstiges:	Nachgewiesen in: MEDLINE Sprachen: English Publication Type: Journal Article Language: English [Fish Shellfish Immunol] 2019 Oct; Vol. 93, pp. 90-98. <i>Date of Electronic Publication: </i>2019 Jul 18. MeSH Terms: Fish Diseases / immunology ; Fishes / genetics ; Fishes / immunology ; Gene Expression Regulation / immunology ; Immunity, Innate / genetics ; Interferon Regulatory Factor-2 / genetics ; Interferon Regulatory Factor-2 / *immunology ; Amino Acid Sequence ; Animals ; Base Sequence ; Female ; Fish Proteins / chemistry ; Fish Proteins / genetics ; Fish Proteins / immunology ; Flagellin / pharmacology ; Gene Expression Profiling / veterinary ; Interferon Regulatory Factor-2 / chemistry ; Lipopolysaccharides / pharmacology ; Male ; Phylogeny ; Poly I-C / pharmacology ; Promoter Regions, Genetic Contributed Indexing: Keywords: IFNa3; IRF2; Promoter activity; Trachinotus ovatus; Transcription factors Substance Nomenclature: 0 (Fish Proteins) ; 0 (Interferon Regulatory Factor-2) ; 0 (Lipopolysaccharides) ; 12777-81-0 (Flagellin) ; O84C90HH2L (Poly I-C) Entry Date(s): Date Created: 20190722 Date Completed: 20200106 Latest Revision: 20200106 Update Code: 20240513

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Functional characterization of interferon regulatory factor 2 and its role in the transcription of interferon a3 in golden pompano Trachinotus ovatus (Linnaeus 1758).