Gilardi P, Perricaudet M
Nucleic Acids Res. 1986 Nov 25;14(22):9035-49. doi: 10.1093/nar/14.22.9035.
To study how the E1A polypeptides of adenovirus type 2 regulate transcription, we have constructed chimeric plasmids containing the bacterial gene encoding chloramphenicol acetyl transferase (CAT) under the control of either the wild type or the deleted E4 promoter of adenovirus type 2. Our previous results showed that promoter sequences located upstream from position -158, as measured from the cap site, are essential to the transactivation process. From a new set of deletion mutants, we now show that two regions, located between positions -239 and -218 and between positions -179 and -158, are involved in the E1A transactivation process. The deletion of only one of them does not significantly alter the E1A induction process compared with the wild type. Moreover, we show that these two regions lie within a DNA fragment which possesses the properties of an E1A-inducible "enhancer-like" element. In addition, the DNA fragment which contains this enhancer element is also able to confer the E1A inducibility to a heterologous promoter.
为了研究2型腺病毒的E1A多肽如何调节转录,我们构建了嵌合质粒,这些质粒含有在2型腺病毒野生型或缺失的E4启动子控制下编码氯霉素乙酰转移酶(CAT)的细菌基因。我们之前的结果表明,从帽位点开始测量,位于-158位上游的启动子序列对于反式激活过程至关重要。从一组新的缺失突变体中,我们现在表明,位于-239至-218位之间以及-179至-158位之间的两个区域参与了E1A反式激活过程。与野生型相比,仅缺失其中一个区域并不会显著改变E1A诱导过程。此外,我们表明这两个区域位于一个具有E1A诱导型“增强子样”元件特性的DNA片段内。此外,包含该增强子元件的DNA片段也能够赋予异源启动子E1A诱导性。