Karls R K, Wolf J R, Donohue T J
Department of Bacteriology, University of Wisconsin-Madison, 312 E. B. Fred Hall, 1550 Linden Drive, Madison, WI 53706, USA.
Mol Microbiol. 1999 Nov;34(4):822-35. doi: 10.1046/j.1365-2958.1999.01649.x.
The Rhodobacter sphaeroides photosynthesis response regulator, PrrA, positively regulates cycA P2 expression. Deletion analysis has identified sequences within 73 bp upstream of the transcription initiation site that are required for the activation of cycA P2 by PrrA. A mutant form of the Rhodobacter capsulatus PrrA homologue, whose activity is independent of phosphorylation (RegA*), protects an approximately 26 bp region of cycA P2 that is centred at approximately -50 from DNase digestion, and activates transcription of a mutant -14T promoter with increased activity when using either R. sphaeroides RNA polymerase or Escherichia coli Esigma70. A 4 bp target site mutation that eliminated DNA binding and transcription activation by RegA* in vitro also abolished PrrA activation of cycA P2 transcription in vivo, indicating that this region contains a PrrA binding site. By analysing the behaviour of the -14T mutant cycA P2 promoter in vivo, we also found that PrrA uses the same target site to activate expression in both the presence and the absence of O2. However, the extent of transcription activation by PrrA at cycA P2 in vivo is greater under anaerobic conditions.
球形红杆菌光合作用响应调节因子PrrA正向调节cycA P2的表达。缺失分析已确定转录起始位点上游73 bp内的序列是PrrA激活cycA P2所必需的。荚膜红杆菌PrrA同源物的一种突变形式,其活性不依赖于磷酸化(RegA*),可保护cycA P2中一个约26 bp的区域,该区域以距DNase消化约-50为中心,并在使用球形红杆菌RNA聚合酶或大肠杆菌Esigma70时激活活性增加的突变-14T启动子的转录。一个4 bp的靶位点突变在体外消除了RegA*的DNA结合和转录激活,在体内也消除了PrrA对cycA P2转录的激活,表明该区域包含一个PrrA结合位点。通过分析-14T突变cycA P2启动子在体内的行为,我们还发现PrrA在有氧和无氧条件下都使用相同的靶位点来激活表达。然而,在体内厌氧条件下,PrrA对cycA P2的转录激活程度更大。