College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China.
Dev Comp Immunol. 2012 Oct;38(2):377-82. doi: 10.1016/j.dci.2012.06.001. Epub 2012 Jun 12.
Interferon regulatory factor 5 (IRF5) has been identified as a key transcriptional mediator regulating expression of both type I interferons (IFNs) and proinflammatory cytokines. In this study, the cDNA and genomic sequences of IRF5 were isolated from Japanese flounder, Paralichthys olivaceus. The gene of Japanese flounder (Jf)IRF5 is 7326 bp long, contains 9 exons and 8 introns and encodes a putative protein of 472 amino acids. The predicted protein sequence shares 61.1-81.9% identity to fish IRF5 and possesses a DNA-binding domain (DBD), a middle region (MR), an IRF association domain (IAD), a virus activated domain (VAD) and two nuclear localization signals (NLSs) conserved in all known IRF5s. Phylogenetic analysis clustered it into the teleost IRF5 subgroup within vertebrate IRF5 group. JfIRF5 mRNA was constitutively expressed in all tissues examined, with higher levels observed in the gills and head kidney. Gene expression of JfIRF5 was analyzed over a 7-day time course in the gills, head kidney, spleen and muscle of Japanese flounders challenged with lymphocystis disease virus (LCDV) and polyinosinic:polycytidylic acid (poly I:C). The data showed that JfIRF5 expression was slightly up-regulated by LCDV, but its induction time was clearly moved up; in contrast, the induction upon poly I:C challenge started not earlier than day 2 post-injection and was stronger and more persistent with a later peak time in all four organs. The late and long-lasting inductive expression of JfIRF5 following poly I:C challenge suggests that it might be an interferon stimulated gene (ISG), the induction of which is driven by poly I:C-induced type I IFNs.
干扰素调节因子 5(IRF5)已被确定为调节 I 型干扰素(IFN)和促炎细胞因子表达的关键转录调节剂。本研究从牙鲆(Paralichthys olivaceus)中分离出 IRF5 的 cDNA 和基因组序列。牙鲆 IRF5 的基因长 7326bp,包含 9 个外显子和 8 个内含子,编码一个推定的 472 个氨基酸的蛋白质。预测的蛋白质序列与鱼类 IRF5 具有 61.1-81.9%的同源性,并且具有 DNA 结合域(DBD)、中间区(MR)、IRF 结合域(IAD)、病毒激活域(VAD)和两个核定位信号(NLS)在所有已知的 IRF5 中保守。系统进化分析将其聚类为脊椎动物 IRF5 组内的硬骨鱼 IRF5 亚组。JfIRF5mRNA 在所有检查的组织中均持续表达,在鳃和头肾中表达水平较高。在牙鲆鳃、头肾、脾脏和肌肉中,用淋巴囊肿病病毒(LCDV)和聚肌苷酸:聚胞苷酸(poly I:C)对 JfIRF5 的基因表达进行了 7 天时间过程的分析。结果表明,LCDV 轻微上调 JfIRF5 的表达,但诱导时间明显提前;相反,poly I:C 刺激后诱导作用不早于注射后第 2 天,在所有四个器官中诱导作用更强、更持久,峰值时间更晚。poly I:C 刺激后 JfIRF5 的晚期和持久诱导表达表明,它可能是干扰素刺激基因(ISG),其诱导作用由 poly I:C 诱导的 I 型 IFN 驱动。