Yoshikawa T, Shimano H, Amemiya-Kudo M, Yahagi N, Hasty A H, Matsuzaka T, Okazaki H, Tamura Y, Iizuka Y, Ohashi K, Osuga J, Harada K, Gotoda T, Kimura S, Ishibashi S, Yamada N
Department of Metabolic Diseases, University of Tokyo, Bunkyo-ku, Tokyo 113-8655, Japan.
Mol Cell Biol. 2001 May;21(9):2991-3000. doi: 10.1128/MCB.21.9.2991-3000.2001.
In an attempt to identify transcription factors which activate sterol-regulatory element-binding protein 1c (SREBP-1c) transcription, we screened an expression cDNA library from adipose tissue of SREBP-1 knockout mice using a reporter gene containing the 2.6-kb mouse SREBP-1 gene promoter. We cloned and identified the oxysterol receptors liver X receptor (LXRalpha) and LXRbeta as strong activators of the mouse SREBP-1c promoter. In the transfection studies, expression of either LXRalpha or -beta activated the SREBP-1c promoter-luciferase gene in a dose-dependent manner. Deletion and mutation studies, as well as gel mobility shift assays, located an LXR response element complex consisting of two new LXR-binding motifs which showed high similarity to an LXR response element recently found in the ABC1 gene promoter, a reverse cholesterol transporter. Addition of an LXR ligand, 22(R)-hydroxycholesterol, increased the promoter activity. Coexpression of retinoid X receptor (RXR), a heterodimeric partner, and its ligand 9-cis-retinoic acid also synergistically activated the SREBP-1c promoter. In HepG2 cells, SREBP-1c mRNA and precursor protein levels were induced by treatment with 22(R)-hydroxycholesterol and 9-cis-retinoic acid, confirming that endogenous LXR-RXR activation can induce endogenous SREBP-1c expression. The activation of SREBP-1c by LXR is associated with a slight increase in nuclear SREBP-1c, resulting in activation of the gene for fatty acid synthase, one of its downstream genes, as measured by the luciferase assay. These data demonstrate that LXR-RXR can modify the expression of genes for lipogenic enzymes by regulating SREBP-1c expression, providing a novel link between fatty acid and cholesterol metabolism.
为了鉴定激活固醇调节元件结合蛋白1c(SREBP-1c)转录的转录因子,我们使用含有2.6 kb小鼠SREBP-1基因启动子的报告基因,筛选了来自SREBP-1基因敲除小鼠脂肪组织的表达cDNA文库。我们克隆并鉴定出氧甾醇受体肝X受体(LXRα)和LXRβ是小鼠SREBP-1c启动子的强激活剂。在转染研究中,LXRα或LXRβ的表达以剂量依赖的方式激活SREBP-1c启动子-荧光素酶基因。缺失和突变研究以及凝胶迁移率变动分析确定了一个由两个新的LXR结合基序组成的LXR反应元件复合体,该复合体与最近在逆向胆固醇转运蛋白ABC1基因启动子中发现的LXR反应元件高度相似。添加LXR配体22(R)-羟基胆固醇可增加启动子活性。视黄酸X受体(RXR)作为异源二聚体伴侣及其配体9-顺式视黄酸的共表达也协同激活SREBP-1c启动子。在HepG2细胞中,用22(R)-羟基胆固醇和9-顺式视黄酸处理可诱导SREBP-1c mRNA和前体蛋白水平升高,证实内源性LXR-RXR激活可诱导内源性SREBP-1c表达。LXR对SREBP-1c的激活与核内SREBP-1c略有增加有关,导致其下游基因之一脂肪酸合酶的基因激活,这通过荧光素酶测定来衡量。这些数据表明,LXR-RXR可通过调节SREBP-1c的表达来改变脂肪生成酶基因的表达,为脂肪酸和胆固醇代谢之间提供了新的联系。