The downstream core promoter element, DPE, is conserved from Drosophila to humans and is recognized by TAF(II)60 of Drosophila
The downstream core promoter element, DPE, is conserved from Drosophila to humans and is recognized by TAF(II)60 of Drosophila
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DOI：
10.1101/gad.11.22.3020
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发表时间：
1997-11-15
影响因子：
10.5
通讯作者：
Kadonaga, JT
中科院分区：
文献类型：
--
作者：
Burke, TW;Kadonaga, JT
我们分析了下游启动子元件（DPE）的功能，这是一个独特的7个核苷酸的核心启动子元件，类似于果蝇中许多TATA盒缺陷（TATA-少）启动子的转录起始位点下游的30个核苷酸。Inr和DPE基序之间的间距有严格的要求，因为Inr和DPE之间的距离增加或减少3个核苷酸会导致转录减少7至8倍，以及纯化的TFIID结合显著减少。这些结果表明TFIID与Inr和DPE序列的特异性和有点刚性的相互作用。用含有TATA较少的DPE的启动子对纯化的TFIID进行光交联分析，揭示了dTAF（II）60和dTAF（II）40与DPE的特异性交联，与dTAF（II）60的交联效率高于与dTAF（II）40的交联效率。这些数据，结合先前充分表征的两个TAF之间的相互作用及其与组蛋白H4和H3的同源性，表明dTAF（II）60-dTAF（II）40异源四聚体与DPE结合。人类和果蝇的转录因子对DPE序列和Inr-DPE间隔的要求基本相同。此外，人干扰素调节因子-1（IRF-1）基因的TATA-少启动子含有一个DPE，该DPE对体外和培养细胞中的转录活性都很重要。因此，这些研究提供了TAF在含TATA-少DPE基因的基础转录中的直接作用的证据，并且共同表明DPE在许多方面是存在于果蝇到人类中的TATA盒的下游对应物。
We analyzed the function of the downstream promoter element (DPE), a distinct 7-nucleotide core promoter element that is similar to 30 nucleotides downstream of the transcription start site of many TATA-box-deficient (TATA-less) promoters in Drosophila. There is a strict requirement for spacing between the Inr and DPE motifs, as an increase or decrease of 3 nucleotides in the distance between the Inr and DPE causes a seven-to eightfold reduction in transcription as well as a significant reduction in the binding of purified TFIID. These results suggest a specific and somewhat rigid interaction of TFIID with the Inr and DPE sequences. Photo-cross-linking analysis of purified TFIID with a TATA-less DPE-containing promoter revealed specific cross-linking of dTAF(II)60 and dTAF(II)40 to the DPE, with a higher efficiency of cross-linking to dTAF(II)60 than to dTAF(II)40. These data, combined with the previously well-characterized interactions between the two TAFs and their homology to histones H4 and H3, suggest that a dTAF(II)60-dTAF(II)40 heterotetramer binds to the DPE. Human and Drosophila transcription factors exhibit essentially the same requirements for DPE sequence and for Inr-DPE spacing. In addition, the TATA-less promoter of the human interferon regulatory factor-1 (IRF-1) gene contains a DPE that is important for transcriptional activity both in vitro and in cultured cells. Hence, these studies provide evidence for a direct role of TAFs in basal transcription of TATA-less DPE-containing genes and collectively indicate that the DPE is, in many respects, a downstream counterpart to the TATA box that is present in Drosophila to humans.