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

在啮齿动物中，高果糖饮食会诱导与代谢综合征类似的代谢进化。以前，我们提出在小鼠肝脏中，统一的核蛋白在-468左右通过SREBP-1 C启动子与区域结合，我们发现单个核肽多态性（SNP）在对高果糖饮食的反应中起着关键作用。在这里，使用MALDI-TOF质量技术，我们确定了X染色体相关的RNA结合基序蛋白（RBMX）作为一种新的候选分子。在EMSA中，抗RBMX抗体置换了果糖喂养诱导的带。 RBMX对大鼠血肿细胞的过表达或抑制调节SREBP-1C启动子活性。 RBMX可以响应高果糖饮食而控制小鼠肝脏中的SREBP-1C表达。Furthermore。阐明蛋白质RBMX在-468在-468的影响下激活SREBP-1C启动子的机制，通过SREBP-1C启动子进行-468的影响，我们对YEAST ONE EASTER INDERIDER IDERIDER IDERIDER IDERES IDERITY IDERES INEERIED IDERES INEERIES INEERIES INEERIES IDESTER IDESTER SEPTOHR INSEERE SELITHES and BEYSES SEFITHES和TRID HYBRID分析，并鉴定LOC673353。 LOC673353的核苷酸序列几乎与18S核糖体RNA基因的一部分相同，并表示为小鼠肝脏中的mRNA和蛋白质。 LOC673353（而不是RBMX）直接与SREBP-1C基因的启动子区域结合，并通过SREBP-1C启动子识别-468的SNP。下拉测定法证明LOC673353与RBMX相互作用。 LOC673353在肝细胞中的过表达可激活SREBP -1C基因的启动子，具体取决于与RBMX的亲和力和-468 bp的SNP。此外，LOC673353的RNA干扰特异性抑制了RBMX诱导的SREBP-1C启动子活性。在小鼠肝脏中，通过使用染色质免疫沉淀测定法检测了LOC673353与SREBP-1C启动子的结合，并通过使用免疫沉淀测定法结合了与RBMX的结合。这些结果清楚地表明，新鉴定的LOC673353通过SREBP-1C启动子直接与-468的SNP区域结合，并通过与RBMX相互作用来调节SREBP-1C启动子。较少的