Wilson K L, Herskowitz I
Proc Natl Acad Sci U S A. 1986 Apr;83(8):2536-40. doi: 10.1073/pnas.83.8.2536.
The STE6 gene of Saccharomyces cerevisiae is an a-specific gene; it is repressed in alpha cells by the alpha 2 product of the mating type locus. To study the role of sequences upstream of STE6 in its regulation and expression, we have determined the DNA sequence of the promoter region, identified the start sites for the STE6 transcript, and identified sequences governing its transcription. Deletions that remove DNA upstream of the STE6 gene were produced and assayed for effects on regulation and expression. The deletions defined two intervals upstream of the STE6 transcription initiation sites. One contains all or part of a negative element; the other contains all or part of a positive element. The negative element is required for repression of STE6 by alpha 2: deletions lacking this element express STE6 constitutively. Such deletions remove a 31-base-pair site, located 135 base pairs upstream of the first transcript start site, that is highly homologous to sites present in the upstream regions of four other genes repressed by alpha 2. These sites are presumably responsible for repression of the a-specific genes by alpha 2. The positive element (a putative upstream activation site) is required for expression of STE6. The deletions define the left boundary of the proposed upstream activation site. Sequence homologies between STE6 and other a-specific genes are found in this region and may mediate activation of this set of genes.
酿酒酵母的STE6基因是一个a型特异性基因;它在α细胞中被交配型基因座的α2产物所抑制。为了研究STE6上游序列在其调控和表达中的作用,我们确定了启动子区域的DNA序列,确定了STE6转录本的起始位点,并确定了控制其转录的序列。产生了去除STE6基因上游DNA的缺失,并检测其对调控和表达的影响。这些缺失定义了STE6转录起始位点上游的两个区间。一个包含全部或部分负调控元件;另一个包含全部或部分正调控元件。负调控元件是α2抑制STE6所必需的:缺乏该元件的缺失会组成型表达STE6。这种缺失去除了一个31个碱基对的位点,该位点位于第一个转录起始位点上游135个碱基对处,与其他四个被α2抑制的基因上游区域中的位点高度同源。这些位点可能负责α2对a型特异性基因的抑制。正调控元件(一个假定的上游激活位点)是STE6表达所必需的。这些缺失定义了所提议的上游激活位点的左边界。在该区域发现了STE6与其他a型特异性基因之间的序列同源性,可能介导了这一组基因的激活。