DNA gyrase is the target for the quinolone drug ciprofloxacin in arabidopsis thaliana

K.M. Evans-Roberts, L.A. Mitchenall, M.K. Wall, Julie Leroux, Josh Mylne, A. Maxwell

Research output: Contribution to journalArticle

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Abstract

© 2016 by The American Society for Biochemistry and Molecular Biology, Inc. The Arabidopsis thaliana genome contains four genes that were originally annotated as potentially encoding DNA gyrase: ATGYRA, ATGYRB1, ATGYRB2, and ATGYRB3. Although we subsequently showed that ATGYRB3 does not encode a gyrase subunit, the other three genes potentially encode subunits of a plant gyrase. We also showed evidence for the existence of supercoiling activity in A. thalianaand that the plantissensitive to quinolone and aminocoumarin antibiotics, compounds that target DNA gyrase in bacteria. However, it was not possible at that time to show whether the A. thaliana genes encoded an active gyrase enzyme, nor whether that enzyme is indeed the target for the quinolone and aminocoumarin antibiotics. Here we show that an A. thaliana mutant resistant to the quinolone drug ciprofloxacin has a point mutation in ATGYRA. Moreover we show that, as in bacteria, the quinolone-sensitive (wild-type) allele is dominant to the resistant gene. Further we have heterologously expressed ATGYRA and ATGYRB2 in a baculovirus expression system and shown supercoiling activity of the partially purified enzyme. Expression/purification of the quin-olone-resistant A. thaliana gyrase yields active enzyme that is resistant to ciprofloxacin. Taken together these experiments now show unequivocally that A. thaliana encodes an organelle-targeted DNA gyrase that is the target of the quinolone drug ciprofloxacin; this has important consequences for plant physiology and the development of herbicides.
Original languageEnglish
Pages (from-to)3136-3144
Number of pages9
JournalJournal of Biological Chemistry
Volume291
Issue number7
Early online date9 Dec 2015
DOIs
Publication statusPublished - 12 Feb 2016

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DNA Gyrase
Quinolones
Ciprofloxacin
Arabidopsis
Genes
Aminocoumarins
Pharmaceutical Preparations
Enzymes
Bacteria
Plant Physiological Phenomena
Anti-Bacterial Agents
Plant Development
Baculoviridae
Physiology
Herbicides
Point Mutation
Organelles
Purification
Alleles
Genome

Cite this

Evans-Roberts, K.M. ; Mitchenall, L.A. ; Wall, M.K. ; Leroux, Julie ; Mylne, Josh ; Maxwell, A. / DNA gyrase is the target for the quinolone drug ciprofloxacin in arabidopsis thaliana. In: Journal of Biological Chemistry. 2016 ; Vol. 291, No. 7. pp. 3136-3144.
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DNA gyrase is the target for the quinolone drug ciprofloxacin in arabidopsis thaliana. / Evans-Roberts, K.M.; Mitchenall, L.A.; Wall, M.K.; Leroux, Julie; Mylne, Josh; Maxwell, A.

In: Journal of Biological Chemistry, Vol. 291, No. 7, 12.02.2016, p. 3136-3144.

Research output: Contribution to journalArticle

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T1 - DNA gyrase is the target for the quinolone drug ciprofloxacin in arabidopsis thaliana

AU - Evans-Roberts, K.M.

AU - Mitchenall, L.A.

AU - Wall, M.K.

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AU - Mylne, Josh

AU - Maxwell, A.

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N2 - © 2016 by The American Society for Biochemistry and Molecular Biology, Inc. The Arabidopsis thaliana genome contains four genes that were originally annotated as potentially encoding DNA gyrase: ATGYRA, ATGYRB1, ATGYRB2, and ATGYRB3. Although we subsequently showed that ATGYRB3 does not encode a gyrase subunit, the other three genes potentially encode subunits of a plant gyrase. We also showed evidence for the existence of supercoiling activity in A. thalianaand that the plantissensitive to quinolone and aminocoumarin antibiotics, compounds that target DNA gyrase in bacteria. However, it was not possible at that time to show whether the A. thaliana genes encoded an active gyrase enzyme, nor whether that enzyme is indeed the target for the quinolone and aminocoumarin antibiotics. Here we show that an A. thaliana mutant resistant to the quinolone drug ciprofloxacin has a point mutation in ATGYRA. Moreover we show that, as in bacteria, the quinolone-sensitive (wild-type) allele is dominant to the resistant gene. Further we have heterologously expressed ATGYRA and ATGYRB2 in a baculovirus expression system and shown supercoiling activity of the partially purified enzyme. Expression/purification of the quin-olone-resistant A. thaliana gyrase yields active enzyme that is resistant to ciprofloxacin. Taken together these experiments now show unequivocally that A. thaliana encodes an organelle-targeted DNA gyrase that is the target of the quinolone drug ciprofloxacin; this has important consequences for plant physiology and the development of herbicides.

AB - © 2016 by The American Society for Biochemistry and Molecular Biology, Inc. The Arabidopsis thaliana genome contains four genes that were originally annotated as potentially encoding DNA gyrase: ATGYRA, ATGYRB1, ATGYRB2, and ATGYRB3. Although we subsequently showed that ATGYRB3 does not encode a gyrase subunit, the other three genes potentially encode subunits of a plant gyrase. We also showed evidence for the existence of supercoiling activity in A. thalianaand that the plantissensitive to quinolone and aminocoumarin antibiotics, compounds that target DNA gyrase in bacteria. However, it was not possible at that time to show whether the A. thaliana genes encoded an active gyrase enzyme, nor whether that enzyme is indeed the target for the quinolone and aminocoumarin antibiotics. Here we show that an A. thaliana mutant resistant to the quinolone drug ciprofloxacin has a point mutation in ATGYRA. Moreover we show that, as in bacteria, the quinolone-sensitive (wild-type) allele is dominant to the resistant gene. Further we have heterologously expressed ATGYRA and ATGYRB2 in a baculovirus expression system and shown supercoiling activity of the partially purified enzyme. Expression/purification of the quin-olone-resistant A. thaliana gyrase yields active enzyme that is resistant to ciprofloxacin. Taken together these experiments now show unequivocally that A. thaliana encodes an organelle-targeted DNA gyrase that is the target of the quinolone drug ciprofloxacin; this has important consequences for plant physiology and the development of herbicides.

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