Asano Atsushi, Jin Hee Kyung, Watanabe Tomomasa
Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan.
Gene. 2003 Mar 13;306:105-13. doi: 10.1016/s0378-1119(03)00428-1.
Mx is an interferon (IFN)-inducible intracellular protein found in various vertebrates that mediates resistance against negative-strand RNA viruses. We have demonstrated previously that feral mouse strains are able to express a functional Mx2 mRNA, while that of the inbred laboratory strains was non-functional because of a single nucleotide insertion in the open reading frame, and under the detectable level. In the present study, we examined the regulation of Mx2 expression in vivo using a congenic mouse carrying Mx1 and Mx2 genes derived from feral strain SPR. Mx2 mRNA was induced strongly in the spleen, ovary and white adipose tissue after the treatment with IFNalpha/beta. Furthermore, we identified the structure of the Mx2 gene. It consists of 14 exons, greatly homologous to the Mx1 gene. The promoter region of Mx2 contained two putative IFN-stimulated response elements (ISREs). We found that the proximal ISRE site positioned between -69 and -55 was essential to IFNalpha/beta-induced transcription by transient transfection assay using reporter gene constructs with mutants of the Mx2 promoter. These results indicate the similarity of the mechanisms of mRNA induction among Mx genes.
Mx是一种在多种脊椎动物中发现的干扰素(IFN)诱导型细胞内蛋白,它介导对负链RNA病毒的抗性。我们之前已经证明,野生小鼠品系能够表达功能性的Mx2 mRNA,而近交系实验室品系的Mx2 mRNA由于开放阅读框中的单个核苷酸插入而无功能,且处于可检测水平之下。在本研究中,我们使用携带源自野生品系SPR的Mx1和Mx2基因的同源小鼠,研究了体内Mx2表达的调控。在用IFNα/β处理后,脾脏、卵巢和白色脂肪组织中的Mx2 mRNA被强烈诱导。此外,我们确定了Mx2基因的结构。它由14个外显子组成,与Mx1基因高度同源。Mx2的启动子区域包含两个假定的干扰素刺激反应元件(ISREs)。我们发现,通过使用带有Mx2启动子突变体的报告基因构建体进行瞬时转染分析,位于-69至-55之间的近端ISRE位点对于IFNα/β诱导的转录至关重要。这些结果表明Mx基因之间mRNA诱导机制的相似性。