Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
J Bacteriol. 2010 Sep;192(17):4327-36. doi: 10.1128/JB.00152-10. Epub 2010 Jun 25.
NikR is a nickel-responsive ribbon-helix-helix transcription factor present in many bacteria and archaea. The DNA binding properties of Escherichia coli and Helicobacter pylori NikR (factors EcNikR and HpNikR, respectively) have revealed variable features of DNA recognition. EcNikR represses a single operon by binding to a perfect inverted repeat sequence, whereas HpNikR binds to promoters from multiple genes that contain poorly conserved inverted repeats. These differences are due in large part to variations in the amino acid sequences of the DNA-contacting beta-sheets, as well as residues preceding the beta-sheets of these two proteins. We present here evidence of another variation in DNA recognition by the NikR protein from Geobacter uraniireducens (GuNikR). GuNikR has an Arg-Gly-Ser beta-sheet that binds specifically to an inverted repeat sequence distinct from those recognized by Ec- or HpNikR. The N-terminal residues that precede the GuNikR beta-sheet residues are required for high-affinity DNA binding. Mutation of individual arm residues dramatically reduced the affinity of GuNikR for specific DNA. Interestingly, GuNikR tetramers are capable of binding cooperatively to the promoter regions of two different genes, nik(MN)1 and nik(MN)2. Cooperativity was not observed for the closely related G. bemidjiensis NikR, which recognizes the same operator sequence. The cooperative mode of DNA binding displayed by GuNikR could affect the sensitivity of transporter gene expression to changes in intracellular nickel levels.
NikR 是一种镍响应性的卷曲-螺旋-环转录因子,存在于许多细菌和古菌中。大肠杆菌和幽门螺杆菌 NikR(分别为 EcNikR 和 HpNikR)的 DNA 结合特性揭示了 DNA 识别的可变特征。EcNikR 通过与完美的反向重复序列结合来抑制单个操纵子,而 HpNikR 则结合到包含不良保守反向重复的多个基因的启动子上。这些差异在很大程度上归因于这两种蛋白质的 DNA 接触β-折叠的氨基酸序列以及β-折叠之前的残基的变化。我们在此展示了来自 Geobacter uraniireducens(GuNikR)的 NikR 蛋白对 DNA 识别的另一种变化的证据。GuNikR 具有 Arg-Gly-Ser β-折叠,该折叠特异性地结合与 Ec-或 HpNikR 识别的不同的反向重复序列。位于 GuNikR β-折叠残基之前的 N 端残基是高亲和力 DNA 结合所必需的。单个臂残基的突变大大降低了 GuNikR 对特定 DNA 的亲和力。有趣的是,GuNikR 四聚体能够协同结合到两个不同基因 nik(MN)1 和 nik(MN)2 的启动子区域。与识别相同操纵子序列的密切相关的 G. bemidjiensis NikR 没有观察到协同作用。GuNikR 显示的协同 DNA 结合模式可能会影响转运蛋白基因表达对细胞内镍水平变化的敏感性。