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TitleCryoEM structures of open dimers of gyrase A in complex with DNA illuminate mechanism of strand passage.
Journal, issue, pagesElife, Vol. 7, Year 2018
Publish dateNov 20, 2018
AuthorsKatarzyna M Soczek / Tim Grant / Peter B Rosenthal / Alfonso Mondragón /
PubMed AbstractGyrase is a unique type IIA topoisomerase that uses ATP hydrolysis to maintain the negatively supercoiled state of bacterial DNA. In order to perform its function, gyrase undergoes a sequence of ...Gyrase is a unique type IIA topoisomerase that uses ATP hydrolysis to maintain the negatively supercoiled state of bacterial DNA. In order to perform its function, gyrase undergoes a sequence of conformational changes that consist of concerted gate openings, DNA cleavage, and DNA strand passage events. Structures where the transported DNA molecule (T-segment) is trapped by the A subunit have not been observed. Here we present the cryoEM structures of two oligomeric complexes of open gyrase A dimers and DNA. The protein subunits in these complexes were solved to 4 Å and 5.2 Å resolution. One of the complexes traps a linear DNA molecule, a putative T-segment, which interacts with the open gyrase A dimers in two states, representing steps either prior to or after passage through the DNA-gate. The structures locate the T-segment in important intermediate conformations of the catalytic cycle and provide insights into gyrase-DNA interactions and mechanism.
External linksElife / PubMed:30457554 / PubMed Central
MethodsEM (single particle)
Resolution4.0 - 6.35 Å
Structure data

9316

6n1p

EMDB-9316, PDB-6n1p:
Dihedral oligomeric complex of GyrA N-terminal fragment with DNA, solved by cryoEM in C2 symmetry
Method: EM (single particle) / Resolution: 6.35 Å

9317

6n1q

EMDB-9317, PDB-6n1q:
Dihedral oligomeric complex of GyrA N-terminal fragment, solved by cryoEM in D2 symmetry
Method: EM (single particle) / Resolution: 5.16 Å

9318

6n1r

EMDB-9318, PDB-6n1r:
Tetrahedral oligomeric complex of GyrA N-terminal fragment, solved by cryoEM in tetrahedral symmetry
Method: EM (single particle) / Resolution: 4.0 Å

Source
  • streptococcus pneumoniae g54 (bacteria)
  • cloning vector pbr322 (others)
KeywordsISOMERASE/DNA / topoisomerase / oligomeric complex / DNA complex / gyrase / T-segment / ISOMERASE-DNA complex / ISOMERASE

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