Functional analysis of differences in transcriptional activity conferred by genetic variants in the 5' flanking region of the IL12RB2 gene.
Functional analysis of differences in transcriptional activity conferred by genetic variants in the 5' flanking region of the IL12RB2 gene.
复制标题
对 IL12RB2 基因 5 侧翼区域的遗传变异所赋予的转录活性差异进行功能分析。
DOI：
10.1007/s00251-015-0882-x
复制
发表时间：
2016
期刊：
影响因子：
3.2
通讯作者：
Nakasho K
中科院分区：
文献类型：
--
作者：
Kato-Kogoe N;Ohyama H;Okano S;Yamanegi K;Yamada N;Hata M;Nishiura H;Abiko Y;Terada N;Nakasho K
白细胞介素12受体β链（IL 12 RB 2）是参与细胞免疫应答的重要调节因子，其基因变异与多种免疫相关疾病的易感性有关。我们先前证明了在IL 12 RB 2的5′侧翼区域具有单核苷酸多态性（SNP）的单倍型，包括− 1035 A>G（rs3762315）和− 1023 A>G（rs3762316），影响IL 12 RB 2的表达，从而改变麻风病和牙周病的易感性。在本研究中，我们确定了与T细胞和NK细胞中IL 12 RB 2单倍型特异性转录活性相关的转录因子。− 1023 G多态性被发现为转录因子激活蛋白（AP）-1创建了一个共有结合位点，基于酶联免疫吸附测定（ELISA）的结合试验表明，这些SNP增强了AP-1与该区域的结合。在报告基因检测中，使用siRNA抑制JunB表达消除了Jurkat T细胞和NK 3.3细胞中含有IL 12 RB 2启动子活性的− 1035 G/− 1023 G和− 1035 A/− 1023 A区域的差异。这些结果表明，−1035/−1023多态性产生了JunB的差异结合亲和力，可能导致IL 12 RB 2的差异表达。此外，− 1035 G和− 1035 A等位基因分别形成了加塔-3和肌细胞增强因子-2（MEF-2）的结合位点。我们的数据表明，除了JunB，-1035/-1023的SNP影响加塔-3和MEF-2的结合亲和力，可能改变IL 12 RB 2的转录活性。这些结果证实了rs3762315和rs3762316对IL 12 RB 2转录的影响。这些遗传变异可能改变T细胞和NK细胞的细胞活化，并改变细胞介导的免疫应答。
Interleukin 12 receptor β chain (IL12RB2) is a crucial regulatory factor involved in cell-mediated immune responses, and genetic variants of the gene encoding IL12RB2 are associated with susceptibility to various immune-related diseases. We previously demonstrated that haplotypes with single nucleotide polymorphisms (SNPs) in the 5′ flanking region ofIL12RB2, including −1035A>G (rs3762315) and −1023A>G (rs3762316), affect the expression of IL12RB2, thereby altering susceptibility to leprosy and periodontal diseases. In the present study, we identified transcription factors associated with the haplotype-specific transcriptional activity ofIL12RB2in T cells and NK cells. The −1023G polymorphism was found to create a consensus binding site for the transcription factor activating protein (AP)-1, and enzyme-linked immunosorbent assay (ELISA)-based binding assays showed that these SNPs enhanced AP-1 binding to this region. In reporter assays, suppression of JunB expression using siRNA eliminated differences in the −1035G/−1023G and −1035A/−1023A regions containingIL12RB2promoter activity in Jurkat T cells and NK3.3 cells. These results suggested that the −1035/−1023 polymorphisms created differential binding affinities for JunB that could lead to differentialIL12RB2expression. Moreover, the −1035G and −1035A alleles formed binding sites for GATA-3 and myocyte enhancer factor-2 (MEF-2), respectively. Our data indicated that in addition to JunB, the SNP at −1035/−1023 influenced GATA-3 and MEF-2 binding affinity, potentially alteringIL12RB2transcriptional activity. These findings confirm the effects of rs3762315 and rs3762316 onIL12RB2transcription. These genetic variants may alter cellular activation of T cells and NK cells and modify cell-mediated immune responses.