A model of EcoRII restriction endonuclease action:: The active complex is most likely formed by one protein subunit and one DNA recognition site
A model of EcoRII restriction endonuclease action:: The active complex is most likely formed by one protein subunit and one DNA recognition site
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DOI：
10.1080/152165499307477
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发表时间：
1999-07-01
期刊：
影响因子：
4.6
通讯作者：
Shabarova, ZA
中科院分区：
文献类型：
--
作者：
Karpova, EA;Kubareva, EA;Shabarova, ZA
为了阐明限制性内切酶EcoR Ⅱ与DNA相互作用的机制，我们通过天然凝胶电泳研究了这种内切酶与一组含有修饰的或典型的识别序列5 ′-d（CCA/TGG）-3 ′的合成DNA-双链体的结合。（i）在与内切核酸酶EcoRII相互作用时，在不存在Mg 2+辅因子的情况下在天然凝胶上形成两种类型的稳定复合物的双链体;（ii）仅形成一种类型的复合物的双链体，在存在和不存在Mg 2+的情况下都观察到，与后者不同，第一组下的双链体可以被内切核酸酶水解，所获得的数据表明，活性复合物最有可能是由一个蛋白质亚基和一个DNA识别序列形成的。EcoRII内切酶的行动模型。
To elucidate the mechanism of interaction of restriction endonuclease EcoRII with DNA, we studied by native gel electrophoresis the binding of this endonuclease to a set of synthetic DNA-duplexes containing the modified or canonical recognition sequence 5'-d(CCA/TGG)-3', All binding substrate or substrate analogues tested could be divided into two major groups: (i) duplexes that, at the interaction with endonuclease EcoRII, form two types of stable complexes on native gel in the absence of Mg2+ cofactor; (ii) duplexes that form only one type of complex, observed both in the presence and absence of Mg2+, Unlike the latter, duplexes under the first group can be hydrolyzed by endonuclease, Data obtained suggest that the active complex is most likely formed by one protein subunit and one DNA recognition sequence. A model of EcoRII endonuclease action is presented.