Yean D, Gralla J
Department of Chemistry & Biochemistry and Molecular Biology Institute, University of California, Los Angeles 90095-1569, USA.
Mol Cell Biol. 1997 Jul;17(7):3809-16. doi: 10.1128/MCB.17.7.3809.
Promoters need to specify both the timing of transcriptional induction and the amount of transcript synthesized. In order to explore each of these effects separately, in vitro assays for the level of active preinitiation complex formation and for the rate of continuous RNA production were done. The effects were found to be influenced differently by different promoter elements. A consensus TATA element had a very strong effect on the rate of continuous RNA production, whereas two types of activators were important primarily in forming active transcription preinitiation complexes. Consensus TATA promoters exhibited high rates of continuous transcription; they assembled active preinitiation transcription complexes slowly but then produced transcripts continuously at an approximately fivefold-higher rate. Initiator-containing TATA-less promoters produced continuous transcripts slowly. Point mutations in the TATA element led to lower levels of transcription by reducing the number of preinitiation complexes and amplifying this reduction by lowering the apparent reinitiation rate. The results allow understanding of the sequence diversity of promoter elements in terms of specifying separate controls over the sensitivity of gene induction and over the strength of the induced promoter.
启动子需要同时确定转录诱导的时间和合成转录本的数量。为了分别探究这些效应,我们针对活性前起始复合物的形成水平和连续RNA产生的速率进行了体外测定。结果发现,不同的启动子元件对这些效应的影响各不相同。共有TATA元件对连续RNA产生的速率有非常强烈的影响,而两类激活因子主要在形成活性转录前起始复合物方面发挥重要作用。共有TATA启动子表现出较高的连续转录速率;它们组装活性前起始转录复合物的速度较慢,但随后以大约五倍的更高速率持续产生转录本。含起始子的无TATA启动子产生连续转录本的速度较慢。TATA元件中的点突变通过减少前起始复合物的数量导致转录水平降低,并通过降低表观重新起始速率放大这种降低。这些结果有助于从对基因诱导敏感性和诱导启动子强度的单独控制方面理解启动子元件的序列多样性。