Cunningham T S, Cooper T G
Department of Microbiology and Immunology, University of Tennessee, Memphis 38163.
J Bacteriol. 1993 Sep;175(18):5851-61. doi: 10.1128/jb.175.18.5851-5861.1993.
Induced expression of the allantoin (DAL) catabolic genes in Saccharomyces cerevisiae has been suggested to be mediated by interaction of three different types of promoter elements. First is an inducer-independent upstream activation sequence, UASNTR, whose operation is sensitive to nitrogen catabolite repression. The GLN3 product is required for UASNTR-mediated transcriptional activation. This site consists of two separated elements, each of which has a GATAA sequence at its core. Response of the DAL genes to inducer is mediated by a second type of cis-acting element, DAL UIS. The DAL82 and DAL81 genes are required for response to inducer; DAL82 protein is the UIS-binding protein. When only the UASNTR and UIS elements are present, DAL gene expression occurs at high levels in the absence of inducer. We, therefore, hypothesized that a third element, an upstream repressor sequence (URS) mediates maintenance of DAL gene expression at a low level when inducer is absent. Since the DAL and UGA genes are overexpressed and largely inducer independent in dal80 deletion mutants, we have suggested DAL80 protein negatively regulates a wide spectrum of nitrogen-catabolic gene expression, likely in conjunction with a URS element. Here we show that DAL80 protein binds to DAL3 and UGA4 upstream DNA sequences, designated URSGATA, consisting of two GATAA-containing sites separated by at least 15 bp. The preferred orientation of the sites is tail to tail, but reasonable binding activity is also observed with a head-to-tail configuration. URSGATA elements contain the sequence GATAA at their core and hence share sequence homology with UASNTR elements.
据推测,酿酒酵母中尿囊素(DAL)分解代谢基因的诱导表达是由三种不同类型的启动子元件相互作用介导的。首先是一个不依赖诱导物的上游激活序列UASNTR,其作用对氮代谢物阻遏敏感。UASNTR介导的转录激活需要GLN3产物。该位点由两个分开的元件组成,每个元件的核心都有一个GATAA序列。DAL基因对诱导物的反应是由第二种顺式作用元件DAL UIS介导的。对诱导物的反应需要DAL82和DAL81基因;DAL82蛋白是UIS结合蛋白。当仅存在UASNTR和UIS元件时,在没有诱导物的情况下DAL基因会高水平表达。因此,我们推测第三种元件,即上游阻遏序列(URS),在没有诱导物时介导DAL基因表达维持在低水平。由于在dal80缺失突变体中DAL和UGA基因过度表达且很大程度上不依赖诱导物,我们认为DAL80蛋白可能与URS元件一起对广泛的氮分解代谢基因表达进行负调控。在此我们表明,DAL80蛋白与DAL3和UGA4上游的DNA序列结合,该序列称为URSGATA,由两个含GATAA的位点组成,它们之间至少相隔15 bp。这些位点的优选方向是尾对尾,但头对头配置也观察到了合理的结合活性。URSGATA元件的核心含有GATAA序列,因此与UASNTR元件具有序列同源性。