Shamblott M J, Leung S, Greene M W, Chen T T
Department of Obstetrics and Gynecology, Division of Developmental Genetics, Johns Hopkins School of Medicine, Baltimore, Maryland 21205, USA.
Mol Mar Biol Biotechnol. 1998 Sep;7(3):181-90.
Insulin-like growth factor I (IGF-I) and IGF-II, acting under the regulatory control of growth hormone, are the principal mediators of vertebrate growth. We have previously demonstrated that like humans, but unlike rodents, rainbow trout maintain high hepatic IGF-II messenger RNA levels into adulthood. Here we describe a rainbow trout IGF-II gene with a proximal promoter that contains two CCAAT/enhancer-binding protein (C/EBP) binding sites (TCBS1 and TCBS2). Nuclear proteins corresponding in size to rat C/EBPalpha and C/EBPbeta were detected on Western immunoblots of growth-hormone-treated and mock-treated trout liver extracts. Electrophoretic mobility shift assay of these nuclear extracts further suggests the presence of C/EBPs in trout liver and confirms the ability of TCBS1 to form a complex with trout liver nuclear proteins that is identical in mobility and specificity to that formed by a mammalian consensus CBS construct. In both Western blot and mobility assay results, the growth-hormone-treated trout livers appeared to have a greater accumulation of C/EBP, suggesting a molecular mechanism by which growth hormone can influence the level of serum IGF-II.
胰岛素样生长因子I(IGF-I)和IGF-II在生长激素的调节控制下起作用,是脊椎动物生长的主要介质。我们之前已经证明,与人类一样,但与啮齿动物不同,虹鳟鱼在成年后肝脏中IGF-II信使核糖核酸水平仍维持在较高水平。在此,我们描述了一个虹鳟鱼IGF-II基因,其近端启动子包含两个CCAAT/增强子结合蛋白(C/EBP)结合位点(TCBS1和TCBS2)。在生长激素处理和模拟处理的鳟鱼肝脏提取物的Western免疫印迹中检测到了大小与大鼠C/EBPα和C/EBPβ相对应的核蛋白。对这些核提取物进行的电泳迁移率变动分析进一步表明鳟鱼肝脏中存在C/EBP,并证实了TCBS1与鳟鱼肝脏核蛋白形成复合物的能力,该复合物在迁移率和特异性上与由哺乳动物共有CBS构建体形成的复合物相同。在Western印迹和迁移率分析结果中,生长激素处理的鳟鱼肝脏似乎积累了更多的C/EBP,这表明生长激素可能通过一种分子机制影响血清IGF-II的水平。