Titlebook: DNA Topoisomerases; Methods and Protocol Duncan J. Clarke Book 2009 Humana Press 2009 Cellular functions.Chromosom.Chromosome structure.DNA

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Cleavage of Plasmid DNA by Eukaryotic Topoisomerase II,I require the enzyme to generate a transient double-stranded break in the backbone of the double helix. To maintain genomic integrity during the cleavage event, topoisomerase II forms covalent bonds between active site tyrosyl residues and the newly generated 5′-DNA termini. In addition to the criti

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Assays for the Preferential Binding of Human Topoisomerase I to Supercoiled DNA,inactive mutant form of the enzyme. In the gel shift assay, the preference for binding to supercoiled plasmid DNA is detected in the presence of linear and nicked forms of the same DNA by a reduction in the mobility of the supercoiled plasmid during electrophoresis in agarose. The more quantitative

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Monitoring the Topoisomerase II DNA Gate Conformational Change with Fluorescence Resonance Energy T in regulating intracellular DNA supercoiling. Topoisomerase II (topo II) performs these DNA transactions by passing one segment of DNA through the other using a reversible, enzyme-bridged double-stranded DNA break. This cleavage/religation of the DNA backbone is coupled to the opening and closing o

Adornment

Single-Molecule Magnetic Tweezers Studies of Type IB Topoisomerases,rticularly suited to the study of topoisomerases due to their unique ability to exert precise and straightforward control of the supercoiled state of DNA. Here, we illustrate in a stepwise fashion how the dynamic properties of type IB topoisomerases can be monitored using this technique.

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