Ogden J E, Stanway C, Kim S, Mellor J, Kingsman A J, Kingsman S M
Mol Cell Biol. 1986 Dec;6(12):4335-43. doi: 10.1128/mcb.6.12.4335-4343.1986.
The Saccharomyces cerevisiae PGK (phosphoglycerate kinase) gene encodes one of the most abundant mRNA and protein species in the cell. To identify the promoter sequences required for the efficient expression of PGK, we undertook a detailed internal deletion analysis of the 5' noncoding region of the gene. Our analysis revealed that PGK has an upstream activation sequence (UASPGK) located between 402 and 479 nucleotides upstream from the initiating ATG sequence which is required for full transcriptional activity. Deletion of this sequence caused a marked reduction in the levels of PGK transcription. We showed that PGK has no requirement for TATA sequences; deletion of one or both potential TATA sequences had no effect on either the levels of PGK expression or the accuracy of transcription initiation. We also showed that the UASPGK functions as efficiently when in the inverted orientation and that it can enhance transcription when placed upstream of a TRP1-IFN fusion gene comprising the promoter of TRP1 fused to the coding region of human interferon alpha-2.
酿酒酵母的磷酸甘油酸激酶(PGK)基因编码细胞中最为丰富的mRNA和蛋白质之一。为了鉴定PGK高效表达所需的启动子序列,我们对该基因5'非编码区进行了详细的内部缺失分析。我们的分析表明,PGK具有一个上游激活序列(UASPGK),位于起始ATG序列上游402至479个核苷酸之间,这是完全转录活性所必需的。删除该序列会导致PGK转录水平显著降低。我们发现PGK对TATA序列没有需求;删除一个或两个潜在的TATA序列对PGK表达水平或转录起始准确性均无影响。我们还表明,UASPGK以反向排列时功能同样高效,并且当置于由TRP1启动子与人类干扰素α-2编码区融合而成的TRP1-IFN融合基因上游时,它能够增强转录。