Leff T, Chambon P
Mol Cell Biol. 1986 Jan;6(1):201-8. doi: 10.1128/mcb.6.1.201-208.1986.
The adenovirus EIa gene products activate transcription from the viral EIII and EIIaE promoters. We studied the mechanism of this stimulation by constructing a series of chimeric promoter recombinants containing the upstream regions of the EIII and EIIaE promoters linked to the TATA box-start-site regions of the viral major late and EIIa late promoters. By introducing these recombinants into HeLa cells together with recombinants producing the EIa gene products, we demonstrated that the induction of EIII and EIIaE transcription by EIa 13S and 12S mRNA products is dependent on sequences located in the upstream region (approximately -40 to -250) of these promoters. In addition, we showed that the major late and EIIa late upstream promoter regions do not contain such EIa-responsive sequence elements. In contrast, after transfection of these chimeric promoter recombinants into 293 cells (which constitutively express the EIa proteins), we found that their relative levels of transcription are similar and markedly different from those observed when they are cotransfected into HeLa cells with EIa protein-producing recombinants. We conclude that the efficiency of transcription from a given promoter in 293 cells is not necessarily related to the presence of a specific EIa-responsive element.
腺病毒E1a基因产物可激活病毒EIII和EIIaE启动子的转录。我们通过构建一系列嵌合启动子重组体来研究这种刺激的机制,这些重组体包含EIII和EIIaE启动子的上游区域,并与病毒主要晚期启动子和EIIa晚期启动子的TATA盒起始位点区域相连。通过将这些重组体与产生E1a基因产物的重组体一起导入HeLa细胞,我们证明E1a 13S和12S mRNA产物对EIII和EIIaE转录的诱导依赖于这些启动子上游区域(约-40至-250)中的序列。此外,我们表明主要晚期和EIIa晚期上游启动子区域不包含此类E1a反应性序列元件。相反,将这些嵌合启动子重组体转染到293细胞(组成性表达E1a蛋白)中后,我们发现它们的相对转录水平相似,且与它们与产生E1a蛋白的重组体共转染到HeLa细胞中时观察到的水平明显不同。我们得出结论,293细胞中给定启动子的转录效率不一定与特定的E1a反应元件的存在有关。