Babiss L E, Herbst R S, Bennett A L, Darnell J E
The Rockefeller University, New York, New York 10021.
Genes Dev. 1987 May;1(3):256-67. doi: 10.1101/gad.1.3.256.
The rat albumin promoter inserted in adenovirus directs transcription in human and rodent hepatoma cells and in rodent hepatocytes (Friedman et al. 1986) and Babiss et al. (1986) but not in HeLa cells or myeloma cells. The nucleotides between -43 and -156 of the RNA start site of the rat albumin gene are required for this cell-specific expression. Protein binding studies (footprints, exonuclease III stops, and gel shifts) all indicate specific interaction in the -80 to -130 region of the gene with factors present in nuclear extracts of hepatocytes and hepatomas, but also from extracts of other cells that do not express the albumin gene. To observe albumin promoter binding, a smaller amount of extract of liver cell nuclei was required compared to extracts of HeLa cell or kidney cell nuclei. In addition, the various tests of DNA-protein interaction did not give qualitatively identical results with extracts from different cells. However, it seems clear that factors are present in several cell types where albumin genes are inactive that will bind to those DNA sequences demonstrated to be necessary for cell-specific expression of this gene. These factors could either be similar but nonidentical factors or the same factors that are modified differently in different cell types.
插入腺病毒的大鼠白蛋白启动子可在人和啮齿动物肝癌细胞以及啮齿动物肝细胞中指导转录(Friedman等人,1986年;Babiss等人,1986年),但在HeLa细胞或骨髓瘤细胞中则不能。大鼠白蛋白基因RNA起始位点-43至-156之间的核苷酸是这种细胞特异性表达所必需的。蛋白质结合研究(足迹法、核酸外切酶III终止法和凝胶迁移率变动分析)均表明,该基因-80至-130区域与肝细胞和肝癌细胞核提取物中的因子存在特异性相互作用,但也与不表达白蛋白基因的其他细胞提取物中的因子存在特异性相互作用。与HeLa细胞核提取物或肾细胞核提取物相比,观察白蛋白启动子结合需要较少的肝细胞细胞核提取物。此外,DNA-蛋白质相互作用的各种测试对不同细胞提取物的结果在性质上并不相同。然而,很明显,在几种白蛋白基因不活跃的细胞类型中存在一些因子,这些因子会与那些已证明对该基因细胞特异性表达必不可少的DNA序列结合。这些因子可能是相似但不同的因子,或者是在不同细胞类型中修饰方式不同的相同因子。