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首页> 外文期刊>African Journal of Biotechnology >RecA-mediated cleavage reaction of Lambda repressor and DNA strand exchange require an active extended filament conformation but not ATP hydrolysis
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RecA-mediated cleavage reaction of Lambda repressor and DNA strand exchange require an active extended filament conformation but not ATP hydrolysis

机译：RecA介导的Lambda阻遏物的切割反应和DNA链交换需要活性的延伸丝构象，但不需要ATP水解

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摘要

DNA pairing and strand exchange activities are essential for genetic recombination. When DNA is damaged, RecA proteins bind to DNA in the presence of ATP and catalyze the specific proteolytic cleavage of Lambda repressor. The cleavage reaction induces and regulates the expression of DNA repair genes. In this work, it has been examined by introducing sites directed mutagenesis (in the ATP catalytic domain or in the DNA binding loop of RecA), the ability of RecA protein to hydrolyze ATP or to cleave Lambda repressor either in the presence of DNA or in the presence of high salt concentration, and the ability of RecA to promote DNA strand exchange. It was observed that mutant E96D does not hydrolyze ATP at all, but fulfills RecA functions such as cleavage of Lambda repressor and strand exchange in the presence of DNA. However mutant E158K hydrolyzes ATP as well in the presence of high salt concentration as in the presence of DNA, but does not fulfill RecA functions. These observations suggest that ATP hydrolysis is not required for the cleavage of Lambda repressor and the genetic recombination, but is necessary for the release of RecA from DNA before DNA repair.
机译：DNA配对和链交换活动对于基因重组至关重要。当DNA受损时，RecA蛋白在ATP存在下与DNA结合并催化Lambda阻遏物的特异性蛋白水解切割。切割反应诱导并调节DNA修复基因的表达。在这项工作中，已通过引入定向诱变位点（在ATP催化域或RecA的DNA结合环中），RecA蛋白在存在DNA或存在DNA的条件下水解ATP或切割Lambda阻遏物的能力进行了研究。高盐浓度的存在以及RecA促进DNA链交换的能力。观察到，突变体E96D根本不水解ATP，但是可以实现RecA功能，例如在DNA存在下切割λ阻遏物和链交换。但是，突变体E158K在高盐浓度下以及在DNA的存在下也能水解ATP，但不能实现RecA功能。这些观察结果表明，ATP水解对于Lambda阻遏物的切割和基因重组不是必需的，但是对于DNA修复前从DNA释放RecA来说是必需的。

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