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Elucidation of the mechanism for the activation of transcription factors and gene response in the aorta of the postprandial state ofrata

阐明餐后状态主动脉转录因子激活和基因反应的机制

基本信息

批准号：
17390262
负责人：
KASHIWAGI Atsunori
金额：
$ 10.6万
依托单位：
Shiga University of Medical Science
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2007
项目状态：
已结题

项目摘要

In rodents a high-fructose diet induces metabolic derangements similar to those in metabolic syndrome. Previously we suggested that in mouse liver an unidentified nuclear protein binding to the region at around -468 by of SREBP-1 c promoter where we found a single nucleotide polymorphism (SNP) plays a key role for the response to high-fructose diet. Here, using MALDI-TOF MASS technique, we identified an X-chromosome-linked RNA binding motif protein (RBMX) as a new candidate molecule. In EMSA, anti-RBMX antibody displaced the bands induced by fructose-feeding. Overexpression or suppression of RBMX on rat hematoma cells regulated the SREBP-1c promoter activity. RBMX may control SREBP-1c expression in mouse liver in response to high-fructose diet.Furthermore, to elucidate the mechanism by which the protein RBMX activates the SREBP-1c promoter under the influence of a SNP at -468 by of SREBP-1c promoter, we performed yeast one-hybrid and two-hybrid analyses and identified the hypothetical … More gene LOC673353. The nucleotide sequence of LOC673353 is almost identical to a part of 18S ribosomal RNA gene and expressed as mRNA and protein in mouse liver. LOC673353, but not RBMX, directly bound to the promoter region of SREBP-1c gene and recognized the SNP at -468 by of SREBP-1c promoter. Pull-down assay demonstrated that LOC673353 interacted with RBMX. The overexpression of LOC673353 in hepatocytes activates the promoter of SREBP-1c gene depending on the affinity to RBMX and the SNP at -468 bp. Furthermore, RNA interference of LOC673353 specifically inhibited the RBMX-induced promoter activity of SREBP-1c. In mouse liver, the binding of LOC673353 to the SREBP-1c promoter was detected by using chromatin immunoprecipitation assay and the binding to RBMX by using the immunoprecipitation assay. These results clearly indicate that the newly identified LOC673353 directly binds to the SNP region at -468 by of SREBP-1c promoter and regulates SREBP-1c promoter through interaction with RBMX. Less

在啮齿类动物中，高果糖饮食诱导代谢紊乱，类似于代谢综合征。以前我们认为，在小鼠肝脏中，一个未鉴定的核蛋白结合到SREBP-1c启动子的-468附近的区域，我们发现该区域的单核苷酸多态性（SNP）在高果糖饮食的反应中起着关键作用。在这里，使用MALDI-TOF MASS技术，我们确定了一个X染色体连锁的RNA结合基序蛋白（RBMX）作为一个新的候选分子。在EMSA中，抗RBMX抗体取代了果糖喂养诱导的条带。RBMX在大鼠血肿细胞上的过表达或抑制调节SREBP-1c启动子活性。此外，为了阐明RBMX蛋白在SREBP-1c启动子-468位点SNP的影响下激活SREBP-1c启动子的机制，我们进行了酵母单杂交和双杂交分析，并确定了假设的RBMX蛋白激活SREBP-1c启动子的机制。 ...更多信息基因LOC 673353。LOC 673353的核苷酸序列与18 S核糖体RNA基因的一部分几乎相同，并在小鼠肝脏中以mRNA和蛋白质形式表达。LOC 673353与SREBP-1c基因启动子区直接结合，识别SREBP-1c启动子区-468位点的SNP，RBMX不与之结合。Pull-down实验表明，LOC 673353与RBMX相互作用。LOC 673353在肝细胞中的过表达依赖于与RBMX的亲和力和-468 bp的SNP激活SREBP-1c基因的启动子。此外，RNA干扰LOC 673353特异性抑制RBMX诱导的SREBP-1c启动子活性。在小鼠肝脏中，LOC 673353与SREBP-1c启动子的结合通过染色质免疫沉淀法检测，与RBMX的结合通过免疫沉淀法检测。这些结果清楚地表明，新鉴定的LOC 673353直接结合到SREBP-1c启动子的-468位点的SNP区域，并通过与RBMX的相互作用调节SREBP-1c启动子。少