Promoter analysis of the interferon regulatory factor 2 and the SNP analysis of the gene

干扰素调节因子2启动子分析及基因SNP分析

基本信息

批准号：
15591187
负责人：
FUKAI Kazuyoshi
金额：
$ 2.24万
依托单位：
Osaka City University Graduate School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591187/
关键词：
IRF2 polymorphism promoter psoriasis 多因子遺伝遺伝子多型転写因子 USF

项目摘要

Interferon regulatory factor 2 (IRF-2) is a transcriptional regulatory protein which represses the expression of interferon-alpha/beta genes. In mice lacking IRF-2, the inflammatory skin disease very similar to human psoriasis develops spontaneously. In addition, IRF-2 gene is located at human chromosome 4q35, where a familial psoriasis susceptibility locus has been mapped. Therefore, we hypothesized the IRF-2 gene is a possible candidate gene for psoriasis.Genotyping of -535G>A was performed by NlaIII-restriction flagment polymorphism (RFLP). Gel-shift and super-shift analysis were employed for the accessment of the binding abilities of transcription factors. To demonstrate differential transcriptional activities, luciferase reporter assays were perforemed.We identified a -535G>A polymorphism in the IRF-2 transcriptional promoter. The -535A allele was more frequent (p= 0.035) in early-onset (less than 41 years of age) psoriasis in Japan, although this was not associated with the old-onset (41 years or older) psoriasis. The -535G allele was found to bind upstream stimulatory factor (USF) by the supershift analysis, whereas the -535A allele did not bind USF. However, the reporter assay revealed no statistical difference of the transcriptional activity between -535A allele and -535G allele. Furthermore, co-transfection of the USF-1 expression vector did not stimulate the promoter activity of our luciferase reporter system.

干扰素调节因子2（IRF-2）是一种转录调节蛋白，可抑制干扰素-α/β基因的表达。在缺乏IRF-2的小鼠中，炎性皮肤病与人牛皮癣非常相似。此外，IRF-2基因位于人类第4q35染色体上，该家族性牛皮癣易感性位点已被映射。因此，我们假设IRF-2基因是牛皮癣的可能候选基因。-535G> a的基型a是通过Nlaiii-Respartictrict straptriction jlagment opyrymormorlist（RFLP）进行的。凝胶变度和超班转移分析用于转录因子的结合能力。为了证明差异转录活性，对荧光素酶报告基因分析进行了穿孔。我们在IRF -2转录启动子中确定了-535G>一种多态性。日本早发（小于41岁）牛皮癣的-535a等位基因在日本的早期（p = 0.035）更为频繁，尽管这与旧式（41岁或41岁）的牛皮癣无关。发现-535G等位基因通过超速移分析结合上游刺激因子（USF），而-535a等位基因没有结合USF。但是，记者测定法显示-535a等位基因和-535G等位基因之间的转录活性没有统计学差异。此外，USF-1表达载体的共转染不会刺激我们的荧光素酶报告基因的启动子活性。