Pagel J M, Winkelman J W, Adams C W, Hatfield G W
Department of Microbiology and Molecular Genetics, College of Medicine, University of California, Irvine 92717.
J Mol Biol. 1992 Apr 20;224(4):919-35. doi: 10.1016/0022-2836(92)90460-2.
It is becoming increasingly clear that the intrinsic and protein-induced topological properties of the DNA helix influence transcriptional efficiency. In this report we describe the properties of two upstream activating regions that influence transcription from the non-overlapping tandem promoters of the ilvGMEDA operon of Escherichia coli. One 20 base-pair region between the promoter sites contains an intrinsic DNA bend that activates transcription from the downstream promoter. The other region contains an integration host factor (IHF) binding site that overlaps the upstream promoter site. IHF binding at this site represses transcription from the upstream promoter and enhances transcription from the downstream promoter. IHF also induces a severe bend in the DNA at its target binding site in the upstream promoter region. The activating property of the 20 base-pair DNA sequence located between the promoters is dependent upon the helical phasing of the sequence-directed DNA bend that it encodes. However, the IHF-mediated activation of transcription is not dependent upon the helical phasing (spatial orientation) of the upstream IHF and downstream promoter sites. The IHF-mediated activation of transcription is also uninfluenced by the presence or absence of the intrinsic DNA bend between its binding site and the downstream promoter site. These results suggest the interesting possibility that IHF activates transcription from the nearby downstream promoter simply by bending the DNA helix in the absence of specific IHF-RNA polymerase or upstream DNA-RNA polymerase interactions.
越来越清楚的是,DNA螺旋的内在拓扑性质和蛋白质诱导的拓扑性质会影响转录效率。在本报告中,我们描述了两个上游激活区域的性质,它们影响大肠杆菌ilvGMEDA操纵子非重叠串联启动子的转录。启动子位点之间的一个20个碱基对的区域包含一个内在的DNA弯曲,可激活下游启动子的转录。另一个区域包含一个与上游启动子位点重叠的整合宿主因子(IHF)结合位点。IHF在此位点的结合会抑制上游启动子的转录,并增强下游启动子的转录。IHF还会在上游启动子区域的靶结合位点诱导DNA发生严重弯曲。启动子之间20个碱基对DNA序列的激活特性取决于其编码的序列导向DNA弯曲的螺旋相位。然而,IHF介导的转录激活并不取决于上游IHF和下游启动子位点的螺旋相位(空间取向)。IHF介导的转录激活也不受其结合位点与下游启动子位点之间是否存在内在DNA弯曲的影响。这些结果提示了一种有趣的可能性,即IHF可能仅通过在不存在特定IHF- RNA聚合酶或上游DNA- RNA聚合酶相互作用的情况下使DNA螺旋弯曲,从而激活附近下游启动子的转录。