Hen R, Borrelli E, Sassone-Corsi P, Chambon P
Nucleic Acids Res. 1983 Dec 20;11(24):8747-60. doi: 10.1093/nar/11.24.8747.
A chimeric recombinant, containing the 270 bp left-terminal fragment of Adenovirus-2 (Ad2) inserted upstream from the -34 to +33 Ad2 major late promoter (Ad2MLP) element, has been used to characterize the transcription stimulatory element which is located at least 231 bp upstream from the E1A capsite in the left-end of Ad2 (Ref. 1). We demonstrate that this element, which acts in cis, possesses several properties characteristic of transcriptional enhancers. Firstly, it potentiates initiation of transcription from the capsite of the heterologous Ad2MLP and from "cryptic" sites often preceded by TATA box-like sequences. Secondly, although there is no critical distance requirement between the enhancer element and the Ad2MLP, the extent of stimulation decreases as the distance between the two element increases. However, in contrast to the other known viral or cellular enhancers which are bidirectional, the Ad2 enhancer is unidirectional, i.e. it potentiates the Ad2MLP element only when it is inserted in its "natural" orientation with respect to the direction of transcription. Using two convergent series of deletions, we have localized the Ad2 enhancer element within a 24 bp segment located at approximately 160 bp from the Ad2 left-end, i.e. 340 bp upstream from the E1A capsite. This 24 bp segment contains a sequence which exhibits a striking homology with the consensus sequence of several viral and cellular enhancers.
一种嵌合重组体已被用于鉴定转录刺激元件,该重组体包含插入到腺病毒2型(Ad2)主要晚期启动子(Ad2MLP)元件-34至+33位点上游的270 bp Ad2左末端片段(参考文献1)。Ad2左末端的E1A帽位点上游至少231 bp处存在该转录刺激元件。我们证明,这种顺式作用元件具有转录增强子的几个特性。首先,它增强了从异源Ad2MLP帽位点以及通常由类TATA盒序列引导的“隐蔽”位点起始转录的能力。其次,尽管增强子元件与Ad2MLP之间没有关键距离要求,但随着这两个元件之间距离的增加,刺激程度会降低。然而,与其他已知的双向病毒或细胞增强子不同,Ad2增强子是单向的,即仅当它相对于转录方向以“自然”方向插入时,它才增强Ad2MLP元件。通过两个收敛的缺失系列,我们已将Ad2增强子元件定位在距Ad2左端约160 bp(即E1A帽位点上游340 bp)的一个24 bp片段内。这个24 bp片段包含一个与几种病毒和细胞增强子的共有序列具有显著同源性的序列。