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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)

文献类型： 外文期刊

作者： Zhu, Ke-Cheng 1 ; Guo, Hua-Yang 1 ; Zhang, Nan 1 ; Guo, Liang 1 ; Liu, Bao-Suo 1 ; Jiang, Shi-Gui 1 ; Zhang, Dian-Chan 1 ;

作者机构： 1.Chinese Acad Fishery Sci, South China Sea Fisheries Res Inst, Minist Agr & Rural Affairs, Key Lab South China Sea Fishery Resources Exploit, Guangzhou 510300, Guangdong, Peoples R China

2.Guangdong Prov Engineer Technol Res Ctr Marine Bi, Guangzhou, Guangdong, Peoples R China

3.Guangdong Prov Key Lab Fishery Ecol & Environm, Guangzhou, Guangdong, Peoples R China

关键词： Trachinotus ovatus; Promoter activity; Transcription factors; IRF2; IFNa3

期刊名称：FISH & SHELLFISH IMMUNOLOGY （ 影响因子：4.581； 五年影响因子：4.851 ）

ISSN： 1050-4648

年卷期： 2019 年 93 卷

页码：

收录情况： SCI

摘要： 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 (To1FNa3), 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 immunerelated tissues (blood, liver, and head-kidney). Moreover, to investigate whether ToIRF2 was a regulator of TolFNa3, 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 TolFNa3 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 TolFNa3 promoter region to regulate TolFNa3 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 TolFNa3 transcription by ToIRF2 and contributes to a better understanding of the transcriptional mechanisms of ToIRF2 in fish.

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