CryoEM structures of open dimers of gyrase A in complex with DNA illuminate mechanism of strand passage
journal contributionposted on 14.09.2020, 10:13 by Katarzyna M Soczek, Tim Grant, Peter B Rosenthal, Alfonso Mondragón
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.
B. subtilisT-segmentcryoEMgyrasemolecular biophysicsstructural biologystructuretopoisomerasesAdenosine TriphosphateBinding SitesCloning, MolecularCryoelectron MicroscopyDNA CleavageDNA GyraseDNA, BacterialEscherichia coliGene ExpressionGenetic VectorsIsoenzymesModels, MolecularOligonucleotidesProtein BindingProtein Conformation, alpha-HelicalProtein Conformation, beta-StrandProtein Interaction Domains and MotifsProtein MultimerizationProtein SubunitsRecombinant ProteinsStreptococcus pneumoniaeRosenthal FC001143SB-ack0601 Biochemistry and Cell Biology