Tiley L S, Madore S J, Malim M H, Cullen B R
Howard Hughes Medical Institute, Duke University Medical Center, Durham, North Carolina 27710.
Genes Dev. 1992 Nov;6(11):2077-87. doi: 10.1101/gad.6.11.2077.
Among eukaryotic transcription trans-activators, the human immunodeficiency virus type 1 (HIV-1) Tat protein is exceptional in that its target site TAR is an RNA rather than a DNA sequence. Here, we confirm that fusion of Tat to the RNA-binding domain of the HIV-1 Rev protein permits the efficient activation of an HIV-1 long terminal repeat (LTR) promoter in which critical TAR sequences have been replaced by RNA sequences derived from the HIV-1 Rev response element (RRE). An RRE target sequence as small as 13 nucleotides is shown to form an effective in vivo target for Rev binding. More important, a fusion protein consisting of Rev attached to the VP16 transcription activation domain was also observed to efficiently activate the HIV-1 LTR from this nascent RNA target. These data demonstrate that trans-activation of transcription by acidic activation domains does not require a stable interaction with the promoter DNA and suggest that VP16, like Tat, can act on steps subsequent to the formation of the HIV-1 LTR preinitiation complex. The finding that the activation domains of VP16 and Tat are functionally interchangeable raises the possibility that these apparently disparate viral trans-activators may nevertheless act via similar mechanisms.
在真核转录反式激活因子中,人类免疫缺陷病毒1型(HIV-1)的Tat蛋白很特殊,因为其靶位点TAR是一个RNA序列而非DNA序列。在此,我们证实将Tat与HIV-1 Rev蛋白的RNA结合结构域融合,能够有效激活HIV-1长末端重复序列(LTR)启动子,其中关键的TAR序列已被源自HIV-1 Rev反应元件(RRE)的RNA序列所取代。一个小至13个核苷酸的RRE靶序列被证明可形成Rev结合的有效体内靶标。更重要的是,还观察到由连接到VP16转录激活结构域的Rev组成的融合蛋白能从这个新生RNA靶标有效激活HIV-1 LTR。这些数据表明,酸性激活结构域对转录的反式激活并不需要与启动子DNA形成稳定的相互作用,并提示VP16与Tat一样,可作用于HIV-1 LTR起始前复合物形成之后的步骤。VP16和Tat的激活结构域在功能上可互换这一发现,增加了这些明显不同的病毒反式激活因子可能通过相似机制发挥作用的可能性。