Li B, Wing H, Lee D, Wu H C, Busby S
School of Biochemistry, The University of Birmingham, Birmingham B15 2TT, UK.
Nucleic Acids Res. 1998 May 1;26(9):2075-81. doi: 10.1093/nar/26.9.2075.
During transcription activation at FNR-dependent promoters where the DNA site for FNR overlaps the -35 element, a surface-exposed activating region in the upstream subunit of the FNR dimer interacts with the C-terminal domain of the RNA polymerase alpha subunit. Starting with a cloned fnr gene encoding a defective FNR derivative carrying substitutions in this activating region, we screened a library of random mutations to identify substitutions that restored FNR activity. Activity can be restored by substitutions at residues T118, E47 and K60. The locations of these residues identify three separate surface-exposed regions of FNR that can play a role in transcription activation. These three regions appear to be analogues of Activating Region 1, Activating Region 2 and Activating Region 3 of the cyclic AMP receptor protein, CRP: our results underscore the similarities between FNR and CRP.
在FNR依赖型启动子的转录激活过程中,FNR的DNA结合位点与-35元件重叠,FNR二聚体上游亚基表面暴露的激活区域与RNA聚合酶α亚基的C末端结构域相互作用。我们从一个克隆的fnr基因开始,该基因编码一种在这个激活区域带有替换的缺陷型FNR衍生物,我们筛选了一个随机突变文库,以鉴定恢复FNR活性的替换。在T118、E47和K60位点的替换可以恢复活性。这些残基的位置确定了FNR的三个独立的表面暴露区域,它们可以在转录激活中发挥作用。这三个区域似乎类似于环磷酸腺苷受体蛋白CRP的激活区域1、激活区域2和激活区域3:我们的结果强调了FNR和CRP之间的相似性。