Yang Xiuqin, Jing Xiaoyan, Song Yanfang, Zhang Caixia, Liu Di
College of Animal Science and Technology, Northeast Agricultural University, Harbin, 150030, China.
Agricultural Academy of Heilongjiang Province, Harbin, 150086, China.
Mol Biol Rep. 2018 Aug;45(4):433-443. doi: 10.1007/s11033-018-4179-7. Epub 2018 Apr 6.
IFN-induced protein with tetratricopeptide repeats 2 (IFIT2) plays important roles in host defense against viral infection as revealed by studies in humans and mice. However, little is known on porcine IFIT2 (pIFIT2). Here, we performed molecular cloning, expression profile, and transcriptional regulation analysis of pIFIT2. pIFIT2 gene, located on chromosome 14, is composed of two exons and have a complete coding sequence of 1407 bp. The encoded polypeptide, 468 aa in length, has three tetratricopeptide repeat motifs. pIFIT2 gene was unevenly distributed in all eleven tissues studied with the most abundance in spleen. Poly(I:C) treatment notably strongly upregulated the mRNA level and promoter activity of pIFIT2 gene. Upstream sequence of 1759 bp from the start codon which was assigned +1 here has promoter activity, and deltaEF1 acts as transcription repressor through binding to sequences at position - 1774 to - 1764. Minimal promoter region exists within nucleotide position - 162 and - 126. Two adjacent interferon-stimulated response elements (ISREs) and two nuclear factor (NF)-κB binding sites were identified within position - 310 and - 126. The ISRE elements act alone and in synergy with the one closer to start codon having more strength, so do the NF-κB binding sites. Synergistic effect was also found between the ISRE and NF-κB binding sites. Additionally, a third ISRE element was identified within position - 1661 to - 1579. These findings will contribute to clarifying the antiviral effect and underlying mechanisms of pIFIT2.
含四肽重复序列的干扰素诱导蛋白2(IFIT2)在人和小鼠的研究中表明,其在宿主抵御病毒感染中发挥重要作用。然而,关于猪IFIT2(pIFIT2)的了解却很少。在此,我们对pIFIT2进行了分子克隆、表达谱及转录调控分析。pIFIT2基因位于14号染色体上,由两个外显子组成,具有1407 bp的完整编码序列。编码的多肽长度为468个氨基酸,具有三个四肽重复基序。pIFIT2基因在所研究的所有11种组织中分布不均,在脾脏中含量最高。聚肌胞苷酸(Poly(I:C))处理显著强烈上调了pIFIT2基因的mRNA水平和启动子活性。从起始密码子(此处定为+1)上游1759 bp的序列具有启动子活性,δEF1通过与-1774至-1764位置的序列结合而作为转录抑制因子。最小启动子区域存在于核苷酸位置-162和-126之间。在-310和-126位置内鉴定出两个相邻的干扰素刺激反应元件(ISRE)和两个核因子(NF)-κB结合位点。ISRE元件单独起作用,并与更靠近起始密码子的元件协同作用,后者具有更强的作用,NF-κB结合位点也是如此。在ISRE和NF-κB结合位点之间也发现了协同效应。此外,在-1661至-1579位置内鉴定出第三个ISRE元件。这些发现将有助于阐明pIFIT2的抗病毒作用及其潜在机制。