de Lorenzo V, Giovannini F, Herrero M, Neilands J B
Department of Biochemistry, University of California, Berkeley 94720.
J Mol Biol. 1988 Oct 20;203(4):875-84. doi: 10.1016/0022-2836(88)90113-1.
Transcription of the iron-controlled aerobactin operon of the enterobacterial plasmid pColV-K30 is negatively regulated through the interaction of a Fe2+-binding repressor (the Fur protein) with operator sequences within the promoter region of the operon. The DNA sequences essential for interaction with the repressor were located by site-directed mutagenesis of specific regions within the 31 base-pair protected by the repressor from DNase I nicking. Occupation of two contiguous repressor-binding sites appears to be required for the complete repression of the system. Contacts of the Fur repressor with the corresponding operator sequences were analyzed with hydroxyl radical footprint and methylation protection experiments. These indicate that DNA-protein contacts approach a symmetrical mode and take place at all sides of the DNA helix.
肠道细菌质粒pColV-K30的铁调控气杆菌素操纵子的转录通过Fe2+结合阻遏物(Fur蛋白)与操纵子启动子区域内的操纵序列相互作用而受到负调控。通过对阻遏物保护免受DNase I切割的31个碱基对特定区域进行定点诱变,确定了与阻遏物相互作用所必需的DNA序列。似乎需要占据两个相邻的阻遏物结合位点才能完全抑制该系统。通过羟基自由基足迹和甲基化保护实验分析了Fur阻遏物与相应操纵序列的接触。这些结果表明,DNA-蛋白质接触接近对称模式,并且发生在DNA螺旋的所有侧面。