Intradomain LexA rotation is a prerequisite for DNA binding specificity
Intradomain LexA rotation is a prerequisite for DNA binding specificity
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DOI：
10.1016/j.febslet.2007.09.006
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发表时间：
2007-10-16
期刊：
影响因子：
3.5
通讯作者：
Zgur-Bertok, Darja
中科院分区：
文献类型：
--
作者：
Butala, Matej;Hodoscek, Milan;Zgur-Bertok, Darja
在没有DNA损伤的情况下，LexA蛋白抑制细菌的SOS系统。我们进行了两种结合到操作符的LexA二聚体的分子动力学模拟。我们的模型预测，LexA DNA结合域相对于二聚c端结构域的旋转是选择性DNA结合所必需的。为了证实该模型，我们设计了双倍和四倍半胱氨酸LexA突变体。电泳迁移分析和表面等离子体共振表明，引入的半胱氨酸残基之间形成的二硫键由于被阻断的结构域重定向而阻止了LexA特异性DNA结合。该模型可为新药设计提供依据。(c) 2007年欧洲生化学会联合会。Elsevier B.V.版权所有。
In the absence of DNA damage the LexA protein represses the bacterial SOS system. We performed molecular dynamic simulations of two LexA dimers bound to operators. Our model predicted that rotation of the LexA DNA binding domain, with respect to the dimerised C-terminal domain, is required for selective DNA binding. To confirm the model, double and quadruple cysteine LexA mutants were engineered. Electrophoretic mobility-shift assay and surface plasmon resonance showed that disulfide bond formation between the introduced cysteine residues precluded LexA specific DNA binding due to blocked domain reorientation. Our model could provide the basis for novel drug design. (c) 2007 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.