Chimeric recombinases with designed DNA sequence recognition
Chimeric recombinases with designed DNA sequence recognition
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DOI：
10.1073/pnas.1533177100
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发表时间：
2003-07-22
影响因子：
11.1
通讯作者：
Stark, WM
中科院分区：
文献类型：
--
作者：
Akopian, A;He, JY;Stark, WM
位点特异性重组通常仅发生在与天然重组酶共同进化的DNA序列之间，以优化序列识别、催化效率和调控。在此，我们表明重组酶的序列识别和催化功能可由不相关的蛋白质结构域来确定。我们描述了一种嵌合重组酶，它具有来自细菌酶Tn3解离酶的一个活化多突变体的催化结构域，该催化结构域与来自小鼠转录因子Zif268的DNA识别结构域相融合。这些蛋白质在由两个Zif268结构域识别的合成靶位点上特异性地催化高效重组。我们的结果证明了解离酶催化结构域的功能自主性，并为创建作用于选定的天然靶序列的“定制”重组酶开辟了道路。
Site-specific recombination typically occurs only between DNA sequences that have co-evolved with a natural recombinase enzyme to optimize sequence recognition, catalytic efficiency, and regulation. Here, we show that the sequence recognition and the catalysis functions of a recombinase can be specified by unrelated protein domains. We describe chimeric recombinases with a catalytic domain from an activated multiple mutant of the bacterial enzyme Tn3 resolvase, fused to a DNA recognition domain from the mouse transcription factor Zif268. These proteins catalyze efficient recombination specifically at synthetic target sites recognized by two Zif268 domains. Our results demonstrate the functional autonomy of the resolvase catalytic domain and open the way to creating "custom-built" recombinases that act at chosen natural target sequences.