Scheper W, Holthuizen P E, Sussenbach J S
Laboratory for Physiological Chemistry, Utrecht University, The Netherlands.
Biochem J. 1996 Aug 15;318 ( Pt 1)(Pt 1):195-201. doi: 10.1042/bj3180195.
Insulin-like growth factor II (IGF-II) is synthesized in many tissues, but the main site of production is the liver. In this paper we show that IGF-II mRNA levels are dependent on the growth conditions of the cells. In Hep3B cells, serum deprivation leads to a marked increase in IGF-II mRNA levels. Serum stimulation of starved Hep3B cells induces a decrease in the amount of IGF-II mRNA, which is not caused by a change in promoter activity. IGF-II mRNAs are subject to endonucleolytic cleavage, a process that requires two widely separated elements in the 3' untranslated region of the mRNA. Specific regions of these elements can form a stable stem structure which is involved in the formation of RNA-protein complexes. By employing electrophoretic mobility shift assays, two complexes have been identified in cytoplasmic extracts of Hep3B cells. The formation of these complexes is related to the growth conditions of the cells and is correlated with the regulation of IGF-II mRNA levels. Our data suggest that, depending on whether serum is present or absent, a transition from one complex to the other occurs. A decrease in the IGF-II mRNA level is also observed when IGF-I or IGF-II is added to serum-deprived Hep3B cells, possibly providing a feedback mechanism for IGF-II production. The serum-induced degradation of IGF-II mRNAs does not require de novo protein synthesis, and is abolished by rapamycin, an inhibitor of p70 S6 kinase.
胰岛素样生长因子II(IGF-II)在许多组织中合成,但主要产生部位是肝脏。在本文中,我们表明IGF-II mRNA水平取决于细胞的生长条件。在Hep3B细胞中,血清剥夺导致IGF-II mRNA水平显著增加。对饥饿的Hep3B细胞进行血清刺激会导致IGF-II mRNA量减少,这不是由启动子活性变化引起的。IGF-II mRNA会发生内切核酸酶切割,这一过程需要mRNA 3'非翻译区中两个相距很远的元件。这些元件的特定区域可形成稳定的茎环结构,该结构参与RNA-蛋白质复合物的形成。通过电泳迁移率变动分析,在Hep3B细胞的细胞质提取物中鉴定出两种复合物。这些复合物的形成与细胞的生长条件有关,并且与IGF-II mRNA水平的调节相关。我们的数据表明,根据血清的存在与否,会发生从一种复合物到另一种复合物的转变。当将IGF-I或IGF-II添加到血清剥夺的Hep3B细胞中时,也观察到IGF-II mRNA水平降低,这可能为IGF-II的产生提供了一种反馈机制。血清诱导的IGF-II mRNA降解不需要从头合成蛋白质,并且被p70 S6激酶的抑制剂雷帕霉素所消除。