Crystallization and initial crystallographic analysis of covalent DNA-cleavage complexes of Staphyloccocus aureus DNA gyrase with QPT-1, moxifloxacin and etoposide

Velupillai Srikannathasan; Alexandre Wohlkonig; Anthony Shillings; Onkar Singh; Pan F Chan; Jianzhong Huang; Michael N Gwynn; Andrew P Fosberry; Paul Homes; Martin Hibbs; Andrew J Theobald; Claus Spitzfaden; Benjamin D Bax

doi:10.1107/S2053230X15015290

Crystallization and initial crystallographic analysis of covalent DNA-cleavage complexes of Staphyloccocus aureus DNA gyrase with QPT-1, moxifloxacin and etoposide

Velupillai Srikannathasan, Alexandre Wohlkonig, Anthony Shillings, Onkar Singh, Pan F Chan, Jianzhong Huang, Michael N Gwynn, Andrew P Fosberry, Paul Homes, Martin Hibbs, Andrew J Theobald, Claus Spitzfaden, Benjamin D Bax

Research output: Contribution to journal › Article › peer-review

21 Citations (Scopus)

Abstract

Fluoroquinolone drugs such as moxifloxacin kill bacteria by stabilizing the normally transient double-stranded DNA breaks created by bacterial type IIA topoisomerases. Previous crystal structures of Staphylococcus aureus DNA gyrase with asymmetric DNAs have had static disorder (with the DNA duplex observed in two orientations related by the pseudo-twofold axis of the complex). Here, 20-base-pair DNA homoduplexes were used to obtain crystals of covalent DNA-cleavage complexes of S. aureus DNA gyrase. Crystals with QPT-1, moxifloxacin or etoposide diffracted to between 2.45 and 3.15 Å resolution. A G/T mismatch introduced at the ends of the DNA duplexes facilitated the crystallization of slightly asymmetric complexes of the inherently flexible DNA-cleavage complexes.

Original language	English
Pages (from-to)	1242-1246
Number of pages	5
Journal	Acta Crystallographica Section F - Structural Biology Communications
Volume	71
Issue number	Pt 10
DOIs	https://doi.org/10.1107/S2053230X15015290
Publication status	Published - 2015

Keywords

Base Sequence
Crystallization
Crystallography, X-Ray
DNA Cleavage
DNA Gyrase/chemistry
Etoposide/chemistry
Fluoroquinolones/chemistry
Heterocyclic Compounds, 4 or More Rings/chemistry
Molecular Sequence Data
Moxifloxacin
Spiro Compounds/chemistry
Staphylococcus aureus/enzymology

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10.1107/S2053230X15015290

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Srikannathasan, V., Wohlkonig, A., Shillings, A., Singh, O., Chan, P. F., Huang, J., Gwynn, M. N., Fosberry, A. P., Homes, P., Hibbs, M., Theobald, A. J., Spitzfaden, C., & Bax, B. D. (2015). Crystallization and initial crystallographic analysis of covalent DNA-cleavage complexes of Staphyloccocus aureus DNA gyrase with QPT-1, moxifloxacin and etoposide. Acta Crystallographica Section F - Structural Biology Communications, 71(Pt 10), 1242-1246. https://doi.org/10.1107/S2053230X15015290

Srikannathasan, Velupillai ; Wohlkonig, Alexandre ; Shillings, Anthony ; Singh, Onkar ; Chan, Pan F ; Huang, Jianzhong ; Gwynn, Michael N ; Fosberry, Andrew P ; Homes, Paul ; Hibbs, Martin ; Theobald, Andrew J ; Spitzfaden, Claus ; Bax, Benjamin D. / Crystallization and initial crystallographic analysis of covalent DNA-cleavage complexes of Staphyloccocus aureus DNA gyrase with QPT-1, moxifloxacin and etoposide. In: Acta Crystallographica Section F - Structural Biology Communications. 2015 ; Vol. 71, No. Pt 10. pp. 1242-1246.

@article{22db32c099c64908a9587283af6f9d72,

title = "Crystallization and initial crystallographic analysis of covalent DNA-cleavage complexes of Staphyloccocus aureus DNA gyrase with QPT-1, moxifloxacin and etoposide",

abstract = "Fluoroquinolone drugs such as moxifloxacin kill bacteria by stabilizing the normally transient double-stranded DNA breaks created by bacterial type IIA topoisomerases. Previous crystal structures of Staphylococcus aureus DNA gyrase with asymmetric DNAs have had static disorder (with the DNA duplex observed in two orientations related by the pseudo-twofold axis of the complex). Here, 20-base-pair DNA homoduplexes were used to obtain crystals of covalent DNA-cleavage complexes of S. aureus DNA gyrase. Crystals with QPT-1, moxifloxacin or etoposide diffracted to between 2.45 and 3.15 {\AA} resolution. A G/T mismatch introduced at the ends of the DNA duplexes facilitated the crystallization of slightly asymmetric complexes of the inherently flexible DNA-cleavage complexes. ",

keywords = "Base Sequence, Crystallization, Crystallography, X-Ray, DNA Cleavage, DNA Gyrase/chemistry, Etoposide/chemistry, Fluoroquinolones/chemistry, Heterocyclic Compounds, 4 or More Rings/chemistry, Molecular Sequence Data, Moxifloxacin, Spiro Compounds/chemistry, Staphylococcus aureus/enzymology",

author = "Velupillai Srikannathasan and Alexandre Wohlkonig and Anthony Shillings and Onkar Singh and Chan, {Pan F} and Jianzhong Huang and Gwynn, {Michael N} and Fosberry, {Andrew P} and Paul Homes and Martin Hibbs and Theobald, {Andrew J} and Claus Spitzfaden and Bax, {Benjamin D}",

year = "2015",

doi = "10.1107/S2053230X15015290",

language = "English",

volume = "71",

pages = "1242--1246",

journal = "Acta Crystallographica Section F - Structural Biology Communications",

issn = "2053-230X",

publisher = "International Union of Crystallography",

number = "Pt 10",

}

Srikannathasan, V, Wohlkonig, A, Shillings, A, Singh, O, Chan, PF, Huang, J, Gwynn, MN, Fosberry, AP, Homes, P, Hibbs, M, Theobald, AJ, Spitzfaden, C & Bax, BD 2015, 'Crystallization and initial crystallographic analysis of covalent DNA-cleavage complexes of Staphyloccocus aureus DNA gyrase with QPT-1, moxifloxacin and etoposide', Acta Crystallographica Section F - Structural Biology Communications, vol. 71, no. Pt 10, pp. 1242-1246. https://doi.org/10.1107/S2053230X15015290

Crystallization and initial crystallographic analysis of covalent DNA-cleavage complexes of Staphyloccocus aureus DNA gyrase with QPT-1, moxifloxacin and etoposide. / Srikannathasan, Velupillai; Wohlkonig, Alexandre; Shillings, Anthony; Singh, Onkar; Chan, Pan F; Huang, Jianzhong; Gwynn, Michael N; Fosberry, Andrew P; Homes, Paul; Hibbs, Martin; Theobald, Andrew J; Spitzfaden, Claus; Bax, Benjamin D.

In: Acta Crystallographica Section F - Structural Biology Communications, Vol. 71, No. Pt 10, 2015, p. 1242-1246.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Crystallization and initial crystallographic analysis of covalent DNA-cleavage complexes of Staphyloccocus aureus DNA gyrase with QPT-1, moxifloxacin and etoposide

AU - Srikannathasan, Velupillai

AU - Wohlkonig, Alexandre

AU - Shillings, Anthony

AU - Singh, Onkar

AU - Chan, Pan F

AU - Huang, Jianzhong

AU - Gwynn, Michael N

AU - Fosberry, Andrew P

AU - Homes, Paul

AU - Hibbs, Martin

AU - Theobald, Andrew J

AU - Spitzfaden, Claus

AU - Bax, Benjamin D

PY - 2015

Y1 - 2015

N2 - Fluoroquinolone drugs such as moxifloxacin kill bacteria by stabilizing the normally transient double-stranded DNA breaks created by bacterial type IIA topoisomerases. Previous crystal structures of Staphylococcus aureus DNA gyrase with asymmetric DNAs have had static disorder (with the DNA duplex observed in two orientations related by the pseudo-twofold axis of the complex). Here, 20-base-pair DNA homoduplexes were used to obtain crystals of covalent DNA-cleavage complexes of S. aureus DNA gyrase. Crystals with QPT-1, moxifloxacin or etoposide diffracted to between 2.45 and 3.15 Å resolution. A G/T mismatch introduced at the ends of the DNA duplexes facilitated the crystallization of slightly asymmetric complexes of the inherently flexible DNA-cleavage complexes.

AB - Fluoroquinolone drugs such as moxifloxacin kill bacteria by stabilizing the normally transient double-stranded DNA breaks created by bacterial type IIA topoisomerases. Previous crystal structures of Staphylococcus aureus DNA gyrase with asymmetric DNAs have had static disorder (with the DNA duplex observed in two orientations related by the pseudo-twofold axis of the complex). Here, 20-base-pair DNA homoduplexes were used to obtain crystals of covalent DNA-cleavage complexes of S. aureus DNA gyrase. Crystals with QPT-1, moxifloxacin or etoposide diffracted to between 2.45 and 3.15 Å resolution. A G/T mismatch introduced at the ends of the DNA duplexes facilitated the crystallization of slightly asymmetric complexes of the inherently flexible DNA-cleavage complexes.

KW - Base Sequence

KW - Crystallization

KW - Crystallography, X-Ray

KW - DNA Cleavage

KW - DNA Gyrase/chemistry

KW - Etoposide/chemistry

KW - Fluoroquinolones/chemistry

KW - Heterocyclic Compounds, 4 or More Rings/chemistry

KW - Molecular Sequence Data

KW - Moxifloxacin

KW - Spiro Compounds/chemistry

KW - Staphylococcus aureus/enzymology

U2 - 10.1107/S2053230X15015290

DO - 10.1107/S2053230X15015290

M3 - Article

C2 - 26457513

VL - 71

SP - 1242

EP - 1246

JO - Acta Crystallographica Section F - Structural Biology Communications

JF - Acta Crystallographica Section F - Structural Biology Communications

SN - 2053-230X

IS - Pt 10

ER -

Srikannathasan V, Wohlkonig A, Shillings A, Singh O, Chan PF, Huang J et al. Crystallization and initial crystallographic analysis of covalent DNA-cleavage complexes of Staphyloccocus aureus DNA gyrase with QPT-1, moxifloxacin and etoposide. Acta Crystallographica Section F - Structural Biology Communications. 2015;71(Pt 10):1242-1246. https://doi.org/10.1107/S2053230X15015290

Crystallization and initial crystallographic analysis of covalent DNA-cleavage complexes of Staphyloccocus aureus DNA gyrase with QPT-1, moxifloxacin and etoposide

Abstract

Keywords

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