PMID:10660569
| Citation |
Mossessova, E, Levine, C, Peng, H, Nurse, P, Bahng, S and Marians, KJ (2000) Mutational analysis of Escherichia coli topoisomerase IV. I. Selection of dominant-negative parE alleles. J. Biol. Chem. 275:4099-103 |
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| Abstract |
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. |
| Links | |
| Keywords |
Alleles; Bacterial Proteins/genetics; DNA Topoisomerase IV; DNA Topoisomerases, Type II/genetics; DNA, Superhelical/chemistry; DNA-Binding Proteins/genetics; Electrophoresis, Polyacrylamide Gel; Escherichia coli/enzymology; Isopropyl Thiogalactoside/metabolism; Microscopy, Fluorescence; Mutation; Phenotype |
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