Herendeen D R, Williams K P, Kassavetis G A, Geiduschek E P
Department of Biology, University of California, San Diego, La Jolla 92093.
Science. 1990 May 4;248(4955):573-8. doi: 10.1126/science.2185541.
Although bacteriophage T4 late promoters are selectively recognized by Escherichia coli RNA polymerase bearing a single protein encoded by T4 gene 55 (gp55), efficient transcription at these promoters requires enhancement by the three T4 DNA polymerase accessory proteins, bound to distal "mobile enhancer" sites. Two principles are shown to govern this transcriptional enhancement: (i) Promoter recognition and communication between the enhancer and the promoter require separate phage-coded proteins. Only RNA polymerase that has the T4 gene 33 protein (gp33) bound to it is subject to enhancement by the three DNA replication proteins. (ii) Transcriptional enhancement in this prokaryotic system is promoter-specific. Promoter specificity is generated by a direct competition of phage T4 gp33 and gp55 with the E. coli promoter recognition protein, sigma 70, for binding to the E. coli RNA polymerase core. Thus, polymerase that contains sigma 70 is competent to transcribe T4 early and middle genes, but lacks the ability to be enhanced by the DNA replication proteins, while polymerase that contains gp55 and gp33 is capable of enhancement via gp33, but its activity is restricted to T4 late promoters by gp55.
尽管噬菌体T4晚期启动子可被携带T4基因55编码的单一蛋白质(gp55)的大肠杆菌RNA聚合酶选择性识别,但在这些启动子处的有效转录需要与远端“移动增强子”位点结合的三种T4 DNA聚合酶辅助蛋白来增强。研究表明有两条原则支配这种转录增强作用:(i)启动子识别以及增强子与启动子之间的通信需要不同的噬菌体编码蛋白。只有结合了T4基因33蛋白(gp33)的RNA聚合酶才能被三种DNA复制蛋白增强。(ii)在这个原核系统中,转录增强具有启动子特异性。启动子特异性是由噬菌体T4 gp33和gp55与大肠杆菌启动子识别蛋白σ70直接竞争结合大肠杆菌RNA聚合酶核心产生的。因此,含有σ70的聚合酶能够转录T4早期和中期基因,但缺乏被DNA复制蛋白增强的能力,而含有gp55和gp33的聚合酶能够通过gp33增强,但它的活性被gp55限制在T4晚期启动子。