Interaction of the DNA topoisomerase II catalytic inhibitor meso-2,3-bis(3,5-dioxopiperazine-1-yl)butane (ICRF-193), a bisdioxopiperazine derivative, with the conserved region(s) of eukaryotic but not prokaryotic enzyme.
ICRF-193 [meso-2,3-bis(3,5-dioxopiperazine-1-yl)butane], a bisdioxopiperazine compound, has been shown to be a catalytic inhibitor of DNA topoisomerase II by stabilizing the enzyme in the form of a closed "protein clamp," an intermediate form in the catalytic cycle (Roca et al., Proc Natl Acad Sci USA 91: 1781-1785, 1994). In view of its usefulness as a probe in the functional analysis of the enzyme, we tried further to define the domain(s) of the enzyme interacting with the drug by examining its inhibitory activity on type II topoisomerases from various species of eukaryotes and prokaryotes. ICRF-193 inhibited the enzyme from yeast, fly, frog, plant, and mammals at IC50 values in the range of 1-13 microM. Experiments using fission yeast truncated mutant type II enzyme lacking both amino-terminal 74 amino acids and carboxy-terminal 265 amino acids revealed that ICRF-193 interacts with the 125 kDa "core" polypeptide of the enzyme. In contrast, prokaryotic type II enzymes, Escherichia coli DNA gyrase, topo IV, and phage T4 topo, were not affected by the drug. From these results, the domain(s) common to eukaryotic but not to prokaryotic type II enzymes interacting with ICRF-193 was speculated.
Sato
M
M
Research Laboratory, Zenyaku Kogyo Co., Ltd., Tokyo, Japan.
Ishida
R
R
Narita
T
T
Kato
J
J
Ikeda
H
H
Fukazawa
H
H
Andoh
T
T
D000595:Amino Acid Sequence; D000818:Animals; D017122:Bacteriophage T4 / Q000201:enzymology; D001665:Binding Sites; D001937:Brassica; D002417:Cattle; D017124:Conserved Sequence; D004250:DNA Topoisomerases, Type II / Q000737:chemistry*; D054659:Diketopiperazines; D004331:Drosophila melanogaster / Q000201:enzymology; D004791:Enzyme Inhibitors / Q000378:metabolism / Q000494:pharmacology; D004926:Escherichia coli / Q000201:enzymology; D005260:Female; D006801:Humans; D008322:Mammals; D010879:Piperazines / Q000378:metabolism / Q000494:pharmacology*; D010920:Placenta / Q000201:enzymology; D010944:Plants / Q000201:enzymology; D010957:Plasmids; D011247:Pregnancy; D012568:Schizosaccharomyces / Q000201:enzymology; D017384:Sequence Deletion; D013045:Species Specificity; D013379:Substrate Specificity; D013950:Thymus Gland / Q000201:enzymology; D059005:Topoisomerase II Inhibitors; D014982:Xenopus laevis
D054659:Diketopiperazines; D004791:Enzyme Inhibitors; D010879:Piperazines; D059005:Topoisomerase II Inhibitors; C070899:4,4'-(1,2-dimethyl-1,2-ethanediyl)bis-2,6-piperazinedione; D004250:DNA Topoisomerases, Type II