The "GyrA-box" is required for the ability of DNA gyrase to wrap DNA and catalyze the supercoiling reaction.
DNA gyrase is the only topoisomerase that can introduce negative supercoils into DNA. It is thought that the binding of conventional type II topoisomerases, including topoisomerase IV, to DNA takes place at the catalytic domain across the DNA gate, whereas DNA gyrase binds to DNA not only at the amino-terminal catalytic domain but also at the carboxyl-terminal domain (CTD) of the GyrA subunit. The binding of the GyrA CTD to DNA allows gyrase to wrap DNA around itself and catalyze the supercoiling reaction. Recent structural studies, however, have revealed striking similarities between the GyrA CTD and the ParC CTD, as well as the ability of the ParC CTD to bind and bend DNA. Thus, the molecular basis of gyrase-mediated wrapping of DNA needs to be reexamined. Here, we have conducted a mutational analysis to determine the role of the "GyrA-box," a 7-amino acid-long motif unique to the GyrA CTD, in determining the DNA binding mode of gyrase. Either a deletion of the entire GyrA-box or substitution of the GyrA-box with 7 Ala residues abolishes the ability of gyrase to wrap DNA around itself and catalyze the supercoiling reaction. However, these mutations do not affect the relaxation and decatenation activities of gyrase. Thus, the presence of a GyrA-box allows gyrase to wrap DNA and catalyze the supercoiling reaction. The consequence of the loss of the GyrA-box during evolution of bacterial type II topoisomerases is discussed.
Kramlinger
Valerie M
VM
Department of Pharmacology, University of Minnesota Medical School-Twin Cities, Minneapolis, Minnesota 55455, USA.
Hiasa
Hiroshi
H
D000255:Adenosine Triphosphate / Q000737:chemistry; D000409:Alanine / Q000737:chemistry; D020816:Amino Acid Motifs; D002384:Catalysis; D020134:Catalytic Domain; D004247:DNA / Q000737:chemistry*; D027081:DNA Gyrase / Q000737:chemistry*; D027101:DNA Topoisomerase IV / Q000737:chemistry; D004269:DNA, Bacterial; D004278:DNA, Superhelical / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology; D009154:Mutation; D009690:Nucleic Acid Conformation; D010802:Phylogeny; D011485:Protein Binding; D014357:Trypsin / Q000494:pharmacology
D004269:DNA, Bacterial; D004278:DNA, Superhelical; D000255:Adenosine Triphosphate; D004247:DNA; D014357:Trypsin; D027101:DNA Topoisomerase IV; D027081:DNA Gyrase; D000409:Alanine