Type II DNA topoisomerase from Saccharomyces cerevisiae is a stable dimer.
Type II DNA topoisomerases function as homodimeric enzymes in transiently cleaving double-stranded DNA to catalyze unlinking and unknotting reactions. The dimeric enzyme creates a DNA double-strand break by forming a covalent attachment between an active site tyrosine from each monomer and a 5'-phosphate from each strand of DNA. The dimer must be very stable to dissociation or subunit exchange when covalently attached to DNA to prevent directly or indirectly catalyzed rearrangements of the genome. Past studies have indicated conflicting results for the monomer-dimer stability of topoisomerase II in solution. Here, we report results from sedimentation equilibrium studies and two different subunit exchange assays indicating that purified Saccharomyces cerevisiae DNA topoisomerase II exists as a stable dimer in solution, with a Kd estimated to be < or = 10(-11) M. This high dimer stability is not detectably altered by a change of ionic strength or by the presence of ATP, ADP, or DNA.
Tennyson
R B
RB
Department of Biochemistry, University of Utah School of Medicine, Salt Lake City 84132, USA.
Lindsley
J E
JE
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000494:pharmacology; D002499:Centrifugation, Density Gradient; D002500:Centrifugation, Isopycnic; D004247:DNA / Q000494:pharmacology; D004250:DNA Topoisomerases, Type II / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D019281:Dimerization; D004795:Enzyme Stability; D008958:Models, Molecular; D009994:Osmolar Concentration; D011487:Protein Conformation; D012441:Saccharomyces cerevisiae / Q000201:enzymology
D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D004247:DNA; D004250:DNA Topoisomerases, Type II