Healy A M, Helser T L, Zitomer R S
Department of Biological Sciences, State University of New York at Albany 12222.
Mol Cell Biol. 1987 Oct;7(10):3785-91. doi: 10.1128/mcb.7.10.3785-3791.1987.
A series of BAL 31 deletions were constructed in the upstream region of the Saccharomyces cerevisiae CYC7 gene to determine sequences required for transcriptional initiation. These deletions identified the TATA box as an alternating A-T sequence at -160 and the initiation sequences as well as the spatial relationship between them. The TATA box was necessary for wild-type levels of expression of the CYC7 gene. Decreasing the distance between the TATA sequence and the initiation site did not alter gene expression, but the site of transcription was shifted 3'-ward. In most cases, transcription initiated at a number of sites, the 5'-most of which was the first suitable site greater than 45 base pairs 3' of the TATA sequence, suggesting a spatial relationship between these sequences. Consensus sequences previously proposed for initiation sites were evaluated with respect to the start sites identified in this study as well as the start sites of other yeast genes.
为确定转录起始所需的序列,在酿酒酵母CYC7基因的上游区域构建了一系列BAL 31缺失片段。这些缺失片段确定了位于-160处的TATA盒为交替的A-T序列、起始序列以及它们之间的空间关系。TATA盒对于CYC7基因野生型表达水平是必需的。减小TATA序列与起始位点之间的距离不会改变基因表达,但转录位点会向3'端移动。在大多数情况下,转录在多个位点起始,其中最5'端的是TATA序列3'端大于45个碱基对的第一个合适位点,这表明这些序列之间存在空间关系。针对本研究中确定的起始位点以及其他酵母基因的起始位点,对先前提出的起始位点共有序列进行了评估。