Effect of casein kinase II-mediated phosphorylation on the catalytic cycle of topoisomerase II. Regulation of enzyme activity by enhancement of ATP hydrolysis.
The catalytic activity of topoisomerase II is stimulated approximately 2-3-fold following phosphorylation by casein kinase II (Ackerman, P., Glover, C. V. C., and Osheroff, N. (1985) Proc. Natl. Acad. Sci. U. S. A. 82, 3164-3168). In order to delineate the mechanism by which the activity of the enzyme is enhanced, the effects of casein kinase II-mediated phosphorylation on the individual steps of the catalytic cycle of Drosophila topoisomerase II were characterized. Phosphorylation did not affect reaction steps that preceded hydrolysis of the enzyme's high energy ATP cofactor. This included enzyme-DNA binding, pre-strand passage DNA cleavage/religation, the double-stranded DNA passage event, and post-strand passage DNA cleavage/religation. In contrast, the rate of topoisomerase II-mediated ATP hydrolysis was stimulated 2.7-fold following phosphorylation by casein kinase II. Since ATP hydrolysis is a prerequisite for enzyme turnover, it is concluded that phosphorylation modulates the overall catalytic activity of topoisomerase II by stimulating the enzyme's ATPase activity.
Corbett
A H
AH
Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-0146.
DeVore
R F
RF
Osheroff
N
N
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D047390:Casein Kinase II; D002384:Catalysis; D004247:DNA / Q000378:metabolism; D004250:DNA Topoisomerases, Type II / Q000378:metabolism*; D004331:Drosophila melanogaster; D006868:Hydrolysis; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*
D000255:Adenosine Triphosphate; D004247:DNA; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases; D004250:DNA Topoisomerases, Type II