Li R, Dickerson N S, Mueller U W, Golden S S
Department of Biology, Texas A&M University, College Station 77843-3258.
J Bacteriol. 1995 Feb;177(3):508-16. doi: 10.1128/jb.177.3.508-516.1995.
The psbAII gene of the cyanobacterium Synechococcus sp. strain PCC 7942 is a member of a three-gene family that encodes the D1 protein of the photosystem II reaction center. Transcription of psbAII is rapidly induced when the light intensity reaching the culture increases from 125 microE.m-2.s-1 (low light) to 750 microE.m-2.s-1 (high light). The DNA segment upstream of psbAII that corresponds to the untranslated leader of its major transcript has enhancer activity and confers high-light induction. We show that one or more soluble proteins from PCC 7942 specifically bind to this region of psbAII (designated the enhancer element). In vivo footprinting showed protein binding to the enhancer element in high-light-exposed cell samples but not in those maintained at low light, even though in vitro mobility shifts were detectable with extracts from low- or high-light-grown cells. When 12 bp were deleted from the psbAII enhancer element, protein binding was impaired and high-light induction of both transcriptional and translational psbAII-lacZ reporters was significantly reduced. This finding indicates that protein binding to this region is required for high-light induction of psbAII. The mutant element also showed impaired enhancer activity when combined with a heterologous promoter.
蓝藻聚球藻属(Synechococcus sp.)菌株PCC 7942的psbAII基因是一个三基因家族的成员,该家族编码光系统II反应中心的D1蛋白。当到达培养物的光强度从125微爱因斯坦·米-2·秒-1(低光)增加到750微爱因斯坦·米-2·秒-1(高光)时,psbAII的转录会迅速被诱导。psbAII上游对应于其主要转录本非翻译前导序列的DNA片段具有增强子活性,并赋予高光诱导性。我们发现来自PCC 7942的一种或多种可溶性蛋白特异性结合到psbAII的这个区域(称为增强子元件)。体内足迹法显示,在高光照射的细胞样品中蛋白质与增强子元件结合,而在低光条件下维持的样品中则不结合,尽管用低光或高光培养的细胞提取物在体外可检测到迁移率变化。当从psbAII增强子元件中缺失12个碱基对时,蛋白质结合受损,psbAII-lacZ转录和翻译报告基因的高光诱导均显著降低。这一发现表明,蛋白质与该区域的结合是psbAII高光诱导所必需的。当与异源启动子结合时,突变元件的增强子活性也受损。