Young M R, Craig E A
McArdle Laboratory for Cancer Research, University of Wisconsin, Madison 53706.
Mol Cell Biol. 1993 Sep;13(9):5637-46. doi: 10.1128/mcb.13.9.5637-5646.1993.
The Saccharomyces cerevisiae HSP70 gene SSA1 has multiple heat shock elements (HSEs). To determine the significance of each of these sequences for expression of SSA1, we analyzed expression from a set of promoters containing point mutations in each of the HSEs, individually and in pairwise combinations. Of the three HSE-like sequences, two (HSE2 and HSE3) were active promoter elements; only one, HSE2, was active under basal growth conditions. Either HSE2 or HSE3 alone was able to drive SSA1 transcription at near-normal rates after heat shock. Both HSE2 and HSE3 were capable of driving basal transcription when placed in the context of the CYC1 promoter. Previous analysis had identified an upstream repressing sequence overlapping HSE2 that repressed basal transcription driven by HSE2. Our analysis showed that basal transcription driven by HSE3 was repressed both by the distant upstream repressing sequence and by closer flanking sequences. The ability to drive basal transcription is not inherent in all natural HSEs, since the HSEs from the heat-inducible SSA3 and SSA4 genes showed no basal activity when placed in the CYC1 vector. Gel mobility shift experiments showed that the same population of heat shock transcription factor molecules bound to HSEs capable of driving basal activity and to HSEs having very low or undetectable basal activity.
酿酒酵母热休克蛋白70基因SSA1有多个热休克元件(HSE)。为了确定这些序列中每一个对SSA1表达的意义,我们分析了一组启动子的表达情况,这些启动子在每个HSE中分别或成对组合含有点突变。在三个类HSE序列中,两个(HSE2和HSE3)是活跃的启动子元件;只有一个,即HSE2,在基础生长条件下是活跃的。热休克后,单独的HSE2或HSE3都能够以接近正常的速率驱动SSA1转录。当置于CYC1启动子的背景下时,HSE2和HSE3都能够驱动基础转录。先前的分析已经确定了一个与HSE2重叠的上游抑制序列,该序列抑制由HSE2驱动的基础转录。我们的分析表明,由HSE3驱动的基础转录受到远处上游抑制序列和更近的侧翼序列的抑制。驱动基础转录的能力并非所有天然HSE所固有,因为来自热诱导型SSA3和SSA4基因的HSE置于CYC1载体中时没有基础活性。凝胶迁移率变动实验表明,相同群体的热休克转录因子分子与能够驱动基础活性的HSE以及基础活性非常低或无法检测到的HSE结合。