EMD-2361
Electron cryo-EM of the full-length Thermus thermophilus DNA gyrase in complex with a 155bp DNA and ciprofloxacin
EMD-2361
Single-particle23.0 Å
Deposition: 13/04/2013Map released: 10/07/2013
Last modified: 18/09/2013
Sample Organism:
Thermus thermophilus,
Escherichia coli
Sample: DNA-bound complex of Thermus thermophilus DNA gyrase with a 155bp DNA in presence of ADPNP and ciprofoxacin.
Deposition Authors: Papillon J, Menetret JF, Batisse C
,
Helye R,
Schultz P ,
Potier N ,
Lamour V
Sample: DNA-bound complex of Thermus thermophilus DNA gyrase with a 155bp DNA in presence of ADPNP and ciprofoxacin.
Deposition Authors: Papillon J, Menetret JF, Batisse C
,
Helye R,
Schultz P ,
Potier N ,
Lamour V
Structural insight into negative DNA supercoiling by DNA gyrase, a bacterial type 2A DNA topoisomerase.
Papillon J,
Menetret JF,
Batisse C ,
Helye R,
Schultz P ,
Potier N ,
Lamour V
(2013) Nucleic Acids Res. , 41 , 7815 - 7827
,
Helye R,
Schultz P ,
Potier N ,
Lamour V
(2013) Nucleic Acids Res. , 41 , 7815 - 7827
Abstract:
Type 2A DNA topoisomerases (Topo2A) remodel DNA topology during replication, transcription and chromosome segregation. These multisubunit enzymes catalyze the transport of a double-stranded DNA through a transient break formed in another duplex. The bacterial DNA gyrase, a target for broad-spectrum antibiotics, is the sole Topo2A enzyme able to introduce negative supercoils. We reveal here for the first time the architecture of the full-length Thermus thermophilus DNA gyrase alone and in a cleavage complex with a 155 bp DNA duplex in the presence of the antibiotic ciprofloxacin, using cryo-electron microscopy. The structural organization of the subunits of the full-length DNA gyrase points to a central role of the ATPase domain acting like a 'crossover trap' that may help to sequester the DNA positive crossover before strand passage. Our structural data unveil how DNA is asymmetrically wrapped around the gyrase-specific C-terminal β-pinwheel domains and guided to introduce negative supercoils through cooperativity between the ATPase and β-pinwheel domains. The overall conformation of the drug-induced DNA binding-cleavage complex also suggests that ciprofloxacin traps a DNA pre-transport conformation.
Type 2A DNA topoisomerases (Topo2A) remodel DNA topology during replication, transcription and chromosome segregation. These multisubunit enzymes catalyze the transport of a double-stranded DNA through a transient break formed in another duplex. The bacterial DNA gyrase, a target for broad-spectrum antibiotics, is the sole Topo2A enzyme able to introduce negative supercoils. We reveal here for the first time the architecture of the full-length Thermus thermophilus DNA gyrase alone and in a cleavage complex with a 155 bp DNA duplex in the presence of the antibiotic ciprofloxacin, using cryo-electron microscopy. The structural organization of the subunits of the full-length DNA gyrase points to a central role of the ATPase domain acting like a 'crossover trap' that may help to sequester the DNA positive crossover before strand passage. Our structural data unveil how DNA is asymmetrically wrapped around the gyrase-specific C-terminal β-pinwheel domains and guided to introduce negative supercoils through cooperativity between the ATPase and β-pinwheel domains. The overall conformation of the drug-induced DNA binding-cleavage complex also suggests that ciprofloxacin traps a DNA pre-transport conformation.