McNeil J B, Dykshoorn P, Huy J N, Small S
Department of Microbiology, University of Toronto, Ontario, Canada.
Curr Genet. 1990 Dec;18(5):405-12. doi: 10.1007/BF00309909.
We show by deletion mutagenesis, followed by in vivo and in vitro analysis, that the binding of a protein factor to the upstream activation sequence (UAS) of the Saccharomyces cerevisiae glycolytic gene PYK, encoding pyruvate kinase, is required for efficient transcription of the corresponding coding region. In addition, gel electrophoretic mobility shift and DNase I protection studies, involving yeast gene products expressed in E. coli, suggest that this trans-acting DNA-binding protein is encoding by the RAP1 gene. The identification of RAP1 binding sites located within the UAS element of the yeast PYK, PGK (phosphoglycerate kinase) and ENO1 (enolase) genes, and in the 5'-upstream region of the ADHI (alcohol dehydrogenase) gene, suggests that a mechanism of coordinate gene expression involving several of the glycolytic genes may exist in yeast.
我们通过缺失诱变，随后进行体内和体外分析表明，一种蛋白质因子与酿酒酵母糖酵解基因PYK（编码丙酮酸激酶）的上游激活序列（UAS）的结合，是相应编码区高效转录所必需的。此外，凝胶电泳迁移率变动和DNase I保护研究，涉及在大肠杆菌中表达的酵母基因产物，表明这种反式作用DNA结合蛋白由RAP1基因编码。在酵母PYK、PGK（磷酸甘油酸激酶）和ENO1（烯醇化酶）基因的UAS元件内以及ADHI（乙醇脱氢酶）基因的5'上游区域中鉴定出RAP1结合位点，这表明酵母中可能存在涉及多个糖酵解基因的协调基因表达机制。
