A Random Screen Using a Novel Reporter Assay System Reveals a Set of Sequences That Are Preferred as the TATA or TATA-Like Elements in the CYC1 Promoter of Saccharomyces cerevisiae.
A Random Screen Using a Novel Reporter Assay System Reveals a Set of Sequences That Are Preferred as the TATA or TATA-Like Elements in the CYC1 Promoter of Saccharomyces cerevisiae.
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DOI：
10.1371/journal.pone.0129357
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Kokubo T
中科院分区：
文献类型：
--
作者：
Watanabe K;Yabe M;Kasahara K;Kokubo T
在酿酒酵母中，II类基因的核心启动子含有TATA或TATA样元件以指导准确的转录起始。全基因组分析表明，TATA元件的共有序列是TATAWAWWR（8 bp），而TATA样元件携带一个或两个错配到这个共识。几个功能不同的TATA序列已被确定的事实表明，这些元件可能发挥作用，至少在某种程度上，在基因特异性的方式。本研究的目的是确定功能TATA序列丰富的一个特定的核心启动子，并比较它们与TATA或TATA样元件，作为前起始复合物（PIC）的装配位点上的酵母基因组。为了这个目的，我们进行了一个随机筛选的TATA元件在CYC1启动子通过使用一种新的报告分析系统，并确定了数百个独特的序列，暂时分为9组。结果表明，7bp TATA元件（即，TATAWAD）和几组TATA样序列是该启动子特别优选的。此外，我们发现，最常见的孤立的TATA样序列，即，TATTTAAA实际上用作内源基因的功能性核心启动子元件，例如，ADE5、7和ADE6。总的来说，这些结果表明，在一个特定的核心启动子中的功能性TATA或TATA样元件的序列要求不是那么严格。然而，这些序列的变异与基因组上的PIC组装位点的变异显著不同，这可能取决于启动子结构并反映了这些序列的基因特异性功能。
In Saccharomyces cerevisiae, the core promoters of class II genes contain either TATA or TATA-like elements to direct accurate transcriptional initiation. Genome-wide analyses show that the consensus sequence of the TATA element is TATAWAWR (8 bp), whereas TATA-like elements carry one or two mismatches to this consensus. The fact that several functionally distinct TATA sequences have been identified indicates that these elements may function, at least to some extent, in a gene-specific manner. The purpose of the present study was to identify functional TATA sequences enriched in one particular core promoter and compare them with the TATA or TATA-like elements that serve as the pre-initiation complex (PIC) assembly sites on the yeast genome. For this purpose, we conducted a randomized screen of the TATA element in the CYC1 promoter by using a novel reporter assay system and identified several hundreds of unique sequences that were tentatively classified into nine groups. The results indicated that the 7 bp TATA element (i.e., TATAWAD) and several sets of TATA-like sequences are preferred specifically by this promoter. Furthermore, we find that the most frequently isolated TATA-like sequence, i.e., TATTTAAA, is actually utilized as a functional core promoter element for the endogenous genes, e.g., ADE5,7 and ADE6. Collectively, these results indicate that the sequence requirements for the functional TATA or TATA-like elements in one particular core promoter are not as stringent. However, the variation of these sequences differs significantly from that of the PIC assembly sites on the genome, presumably depending on promoter structures and reflecting the gene-specific function of these sequences.
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