Positively supercoiled plasmid DNA is produced by treatment of Escherichia coli with DNA gyrase inhibitors

A procedure has been developed whereby the relative amounts of the topoisomers of E. coli plasmid can be determined for cells grown under a variety of conditions. Several applications of the procedure are presented. Addition of either novobiocin or oxolinic acid, two inhibitors of DNA gyrase, gives...

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Bibliographic Details
Published in:Nucleic acids research 1983-05, Vol.11 (10), p.2999-3017
Main Authors: Lockshon, Daniel, Morrs, David R.
Format: Article
Language:English
Subjects:
Aminocoumarins
Coumarins - pharmacology
DNA Replication - drug effects
DNA, Superhelical - genetics
Escherichia coli
Escherichia coli - enzymology
Kinetics
Novobiocin - pharmacology
Nucleic Acid Hybridization
Oxolinic Acid - pharmacology
Plasmids - drug effects
Topoisomerase II Inhibitors
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Summary:A procedure has been developed whereby the relative amounts of the topoisomers of E. coli plasmid can be determined for cells grown under a variety of conditions. Several applications of the procedure are presented. Addition of either novobiocin or oxolinic acid, two inhibitors of DNA gyrase, gives rise to positively supercoiled plasmid. A likely model for the introduction of positive supercoils, involving DNA gyrase itself, is discussed. Qxolinic acid is also shown to induce linearization of plasmid in vivo. Starvation of cells for ATP is shown to cause relaxation of plasmid. The shift of a gyrB temperature-sensitive strain to the restrictive temperature is also shown to cause plasmid relaxation. Finally, it is noted that polyamine starvation of E. coli has no detectable effect on the distribution of topoisomers
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/11.10.2999