Abstract
| Original language | English |
|---|---|
| Pages (from-to) | 3136-3144 |
| Number of pages | 9 |
| Journal | Journal of Biological Chemistry |
| Volume | 291 |
| Issue number | 7 |
| Early online date | 9 Dec 2015 |
| DOIs | |
| Publication status | Published - 12 Feb 2016 |
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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 journal › Article
TY - JOUR
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.
AU - Leroux, Julie
AU - Mylne, Josh
AU - Maxwell, A.
PY - 2016/2/12
Y1 - 2016/2/12
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.
U2 - 10.1074/jbc.M115.689554
DO - 10.1074/jbc.M115.689554
M3 - Article
VL - 291
SP - 3136
EP - 3144
JO - The Journal of Biological Chemistry
JF - The Journal of Biological Chemistry
SN - 0021-9258
IS - 7
ER -