Mutational analysis of Escherichia coli topoisomerase IV.: Selection of dominant-negative parE alleles
Mutational analysis of Escherichia coli topoisomerase IV.: Selection of dominant-negative parE alleles
复制标题
DOI：
10.1074/jbc.275.6.4099
复制
发表时间：
2000-02-11
影响因子：
4.8
通讯作者：
Marians, KJ
中科院分区：
文献类型：
--
作者：
Mossessova, E;Levine, C;Marians, KJ
ParE是拓扑异构酶IV的两个亚基之一，在拓扑异构化过程中参与催化，为了确定ParE的区域，我们开发了一种选择程序来分离显性阴性ParE等位基因。野生型parC和突变型parE都是在中等拷贝质粒上由严格调控的lac启动子表达的，突变的parE等位基因是从导致iptg依赖性细胞死亡的质粒中获救的，当与parC重组形成拓扑异构酶IV时，突变的parE蛋白可以分为两组，一组引发超dna切割，另一组影响共价复合物的形成。
In order to define regions of ParE, one of the two subunits of topoisomerase IV, that are involved in catalysis during topoisomerization, we developed a selection procedure to isolate dominant-negative parE alleles. Both wild-type parC and mutagenized parE were expressed from a tightly-regulated lac promoter on a moderate-copy plasmid, Mutated parE alleles were rescued from those plasmids that caused IPTG-dependent cell death, The mutant ParE proteins could be divided into two groups when reconstituted with ParC to form topoisomerase IV, those that elicited hyper-DNA cleavage and those that affected covalent complex formation.