Friedman A D, Landschulz W H, McKnight S L
Howard Hughes Research Laboratories, Carnegie Institution of Washington, Department of Embryology, Baltimore, Maryland 21210.
Genes Dev. 1989 Sep;3(9):1314-22. doi: 10.1101/gad.3.9.1314.
An expression vector capable of encoding full-length CCAAT/enhancer-binding protein (C/EBP) has been constructed and tested in transient transfection assays for its capacity to activate transcription from the promoter of the serum albumin gene. When tested in cultured hepatoma cells, the C/EBP expression vector achieved potent trans-activation of the albumin promoter. Less substantial activation was observed when the same experiment was conducted using cultured mouse fibroblasts. Expression vectors that encoded defective forms of C/EBP failed to activate the albumin promoter. Moreover, mutated variants of the albumin promoter that lack the C/EBP-binding site failed to be trans-activated. The data are consistent with the interpretation that C/EBP is a bona fide transcription factor. During the course of these experiments it was noted also that C/EBP is more than an order of magnitude less concentrated in cultured hepatoma cells than it is in adult liver cells. Given these findings, we speculate that C/EBP may play a general role in establishing and maintaining the differentiated, nonproliferative state.
一种能够编码全长CCAAT/增强子结合蛋白(C/EBP)的表达载体已构建完成,并在瞬时转染实验中测试了其激活血清白蛋白基因启动子转录的能力。当在培养的肝癌细胞中进行测试时,C/EBP表达载体实现了对白蛋白启动子的高效反式激活。使用培养的小鼠成纤维细胞进行相同实验时,观察到的激活作用较弱。编码缺陷形式C/EBP的表达载体未能激活白蛋白启动子。此外,缺乏C/EBP结合位点的白蛋白启动子突变变体也未能被反式激活。这些数据与C/EBP是一种真正的转录因子这一解释相一致。在这些实验过程中还注意到,与成年肝细胞相比,培养的肝癌细胞中C/EBP的浓度要低一个数量级以上。基于这些发现,我们推测C/EBP可能在建立和维持分化的、非增殖状态中发挥普遍作用。