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Biochemical characterization of ferric uptake regulator (Fur) from Aliivibrio salmonicida. Mapping the DNA sequence specificity through binding studies and structural modelling.
Biometals ( IF 4.1 ) Pub Date : 2020-07-09 , DOI: 10.1007/s10534-020-00240-6
Kristel Berg 1 , Hege Lynum Pedersen 1 , Ingar Leiros 1
Affiliation  

Iron is an essential nutrient for bacteria, however its propensity to form toxic hydroxyl radicals at high intracellular concentrations, requires its acquisition to be tightly regulated. Ferric uptake regulator (Fur) is a metal-dependent DNA-binding protein that acts as a transcriptional regulator in maintaining iron metabolism in bacteria and is a highly interesting target in the design of new antibacterial drugs. Fur mutants have been shown to exhibit decreased virulence in infection models. The protein interacts specifically with DNA at binding sites designated as ‘Fur boxes’. In the present study, we have investigated the interaction between Fur from the fish pathogen Aliivibrio salmonicida (AsFur) and its target DNA using a combination of biochemical and in silico methods. A series of target DNA oligomers were designed based on analyses of Fur boxes from other species, and affinities assessed using electrophoretic mobility shift assay. Binding strengths were interpreted in the context of homology models of AsFur to gain molecular-level insight into binding specificity.



中文翻译:

来自杀鲑鱼的三价铁摄取调节剂 (Fur) 的生化表征。通过结合研究和结构建模绘制 DNA 序列特异性。

铁是细菌的必需营养素,但其在细胞内高浓度下形成有毒羟基自由基的倾向需要对其获取进行严格调控。铁摄取调节剂 (Fur) 是一种金属依赖性 DNA 结合蛋白,在维持细菌中的铁代谢中充当转录调节剂,并且是设计新抗菌药物的一个非常有趣的目标。已显示毛皮突变体在感染模型中表现出降低的毒力。该蛋白质在称为“毛皮盒”的结合位点与 DNA 发生特异性相互作用。在本研究中,我们研究了来自鱼类病原体Aliivibrio Salmonicida 的Fur 之间的相互作用(AsFur) 及其目标 DNA 使用生化和计算机方法的组合。基于对来自其他物种的毛皮盒的分析,以及使用电泳迁移率变化分析评估的亲和力,设计了一系列目标 DNA 寡聚体。在 AsFur 同源模型的背景下解释结合强度,以获得对结合特异性的分子水平洞察。

更新日期:2020-07-09
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