Jansma D B, Archambault J, Mostachfi O, Friesen J D
Department of Genetics, The Hospital for Sick Children, Toronto, Ontario, Canada.
Nucleic Acids Res. 1996 Nov 15;24(22):4543-51. doi: 10.1093/nar/24.22.4543.
We have determined the location of cis-acting elements that are important for the expression of RPO21 and RPO22, genes that encode the two largest subunits of RNA polymerase II (RNAPII) in Saccharomyces cerevisiae. A series of 5'-end deletions and nucleotide substitutions in the upstream regions of RPO21 and RPO22 were tested for their effect on the expression of lacZ fusions of these genes. Deletion of sequences from -723 to -693 in RPO21, which disrupted two Reb1p-binding sites and an Abf1p-binding site, resulted in a 10-fold decrease in expression. A T-rich region downstream of these sites was also important for expression. Deletion of sequences from -437 to -392 in the RPO22-upstream, which resulted in a 30-fold decrease in expression, indicated that the Reb1p- and Abf1p-binding sites in this region were important for RPO22 expression, as was a T-rich sequence immediately downstream of these sites. The RPO21 and RPO22 upstream regions were capable of interacting in vitro (gel-mobility-shift assays) with Reb1p and Abf1p. The similarities in the type and organization of elements in the upstream regions of RPO21 and RPO22 suggest that expression of these genes may be regulated coordinately.
我们已经确定了对酿酒酵母中RNA聚合酶II(RNAPII)两个最大亚基进行编码的基因RPO21和RPO22表达至关重要的顺式作用元件的位置。对RPO21和RPO22上游区域进行了一系列5'端缺失和核苷酸替换,测试它们对这些基因的lacZ融合体表达的影响。RPO21中从-723至-693的序列缺失破坏了两个Reb1p结合位点和一个Abf1p结合位点,导致表达下降10倍。这些位点下游的富含T的区域对表达也很重要。RPO22上游从-437至-392的序列缺失导致表达下降30倍,这表明该区域中的Reb1p和Abf1p结合位点对RPO22表达很重要,这些位点紧邻的富含T的序列也是如此。RPO21和RPO22上游区域能够在体外(凝胶迁移率变动分析)与Reb1p和Abf1p相互作用。RPO21和RPO22上游区域元件的类型和组织方式的相似性表明这些基因的表达可能受到协同调控。