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首页> 外文期刊>Molecular and Cellular Biology >Transcriptional initiation is controlled by upstream GC-box interactions in a TATAA-less promoter.
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Transcriptional initiation is controlled by upstream GC-box interactions in a TATAA-less promoter.

机译：转录起始受无TATAA启动子中上游GC-box相互作用的控制。

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Numerous genes contain TATAA-less promoters, and the control of transcriptional initiation in this important promoter class is not understood. We have determined that protein-DNA interactions at three of the four proximal GC box sequence elements in one such promoter, that of the hamster dihydrofolate reductase gene, control initiation and relative use of the major and minor start sites. Our results indicate that although the GC boxes are apparently equivalent with respect to factor binding, they are not equivalent with respect to function. At least two properly positioned GC boxes were required for initiation of transcription. Abolishment of DNA-protein interaction by site-specific mutation of the most proximal GC box (box I) resulted in a fivefold decrease in transcription from the major initiation site and a threefold increase in heterogeneous transcripts initiating from the vicinity of the minor start site in vitro and in vivo. Mutations that separately abolished interactions at GC boxes II and III while leaving GC box I intact affected the relative utilization of both the major and minor initiation sites as well as transcriptional efficiency of the promoter template in in vitro transcription and transient expression assays. Interaction at GC box IV when the three proximal boxes were in a wild-type configuration had no effect on transcription of the dihydrofolate reductase gene promoter. Thus, GC box interactions not only are required for efficient transcription but also regulate start site utilization in this TATAA-less promoter.
机译：许多基因含有少TATAA的启动子，并且在这种重要的启动子类别中对转录起始的控制尚不清楚。我们已经确定在一个这样的启动子，仓鼠二氢叶酸还原酶基因的四个近端GC框序列元素中的三个，蛋白质-DNA相互作用控制了主要和次要起始位点的起始和相对使用。我们的结果表明，尽管GC盒在因子结合方面看似等效，但在功能上却不等效。至少需要两个正确放置的GC盒才能启动转录。最接近的GC盒（第I盒）的位点特异性突变消除了DNA-蛋白质相互作用，导致从主要起始位点开始的转录转录降低了五倍，而从次要起始位点附近开始的异源转录物增加了三倍。体外和体内。在体外转录和瞬时表达测定中，分别消除了GC框II和III的相互作用而保留GC框I完整的突变影响了主要和次要起始位点的相对利用率以及启动子模板的转录效率。当三个近端盒处于野生型配置时，GC盒IV的相互作用对二氢叶酸还原酶基因启动子的转录没有影响。因此，GC盒相互作用不仅是有效转录所必需的，而且还调节了这种不含TATAA的启动子中起始位点的利用。

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