Structure of the Escherichia coli Fis-DNA complex probed by protein conjugated with 1,10-phenanthroline copper(I) complex.
Structure of the Escherichia coli Fis-DNA complex probed by protein conjugated with 1,10-phenanthroline copper(I) complex.
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通过与 1,10-菲咯啉铜 (I) 复合物缀合的蛋白质探测大肠杆菌 Fis-DNA 复合物的结构。
DOI：
10.1073/pnas.91.5.1721
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发表时间：
1994
影响因子：
11.1
通讯作者：
Johnson,RC
中科院分区：
文献类型：
--
作者：
Pan,CQ;Feng,JA;Finkel,SE;Landgraf,R;Sigman,D;Johnson,RC
大肠杆菌Fis（反转刺激因子）蛋白在许多不同的生物系统中起作用，包括重组、转录和DNA复制。虽然Fis是一种位点特异性的dna结合蛋白，但它缺乏明确的共识识别序列。Fis-DNA复合物的电泳迁移率，以及Fis晶体结构的考虑，表明DNA在Fis结合时发生了显著的变形。为了研究Fis- dna复合物的结构，化学核酸酶1,10-菲罗啉-铜复合物（OP-Cu）被连接到Fis dna结合域内的四个特定位点。其中两种fish - op衍生物在切割DNA方面很活跃。在四个不同的Fis结合位点上获得的断裂模式表明，Fis以非常相似的方式定位在这些高度不同的DNA序列上。Asn-98的衍生物的切割模式通常支持在核心识别序列中包含特定弯曲的fish - dna复合物模型。来自Asn-73的第二个Fis- op衍生物的数据提供了证据，证明Fis蛋白两侧的侧翼DNA比先前假设的更大。侧翼片段的切割效率不同，表明DNA包裹的程度与序列有关。Fis上的特定氨基酸与促进这种DNA包裹有关。
The Escherichia coli Fis (factor for inversion stimulation) protein functions in many diverse biological systems including recombination, transcription, and DNA replication. Although Fis is a site-specific DNA-binding protein, it lacks a well-defined consensus recognition sequence. The electrophoretic mobility of Fis-DNA complexes, along with considerations of the Fis crystal structure, indicates that significant deformation of DNA occurs upon Fis binding. To investigate the structure of Fis-DNA complexes, the chemical nuclease 1,10-phenanthroline-copper complex (OP-Cu) has been linked to four specific sites within the Fis DNA-binding domain. Two of these Fis-OP derivatives were active in cleaving DNA. The scission patterns obtained on four different Fis binding sites indicate that Fis positions itself on these highly divergent DNA sequences in a very similar fashion. The patterns of cleavage of a derivative at Asn-98 generally support a model of a Fis-DNA complex that contains specific bends within the core-recognition sequence. Data from a second Fis-OP derivative at Asn-73 provides evidence for greater wrapping of flanking DNA around the sides of the Fis protein than was previously postulated. The cleavage efficiency of flanking segments varies, suggesting that the extent of DNA wrapping is sequence dependent. Specific amino acids on Fis are implicated in promoting this DNA wrapping.