Local sequence requirements for DNA cleavage by mammalian topoisomerase II in the presence of doxorubicin

Capranico, Giovanni; Kohn, Kurt W.; Pommier, ﻿Yves

doi:10.1093/nar/18.22.6611

Cited by 175 publications

(171 citation statements)

References 27 publications

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“…The molecular details of this binding site are unknown, nevertheless, important information for its location have been revealed by mutational analysis of the cleavage of a preferred site for T4 topoisomerase which suggested that the drugs interact directly with the two bases on either side of the scissile phosphodiester bond (34). Similar conclusions were drawn by studying the local base sequence preference for DNA cleavage by mammalian topoisomerase II in the presence of anti-tumor drugs (35,36). In the case of gyrase, Cove et al (30) found that gyrase shows a preference for a pair of guanine bases being present on either side of the cleaved bond.…”

Section: Discussionmentioning

confidence: 77%

The DNA Gyrase-Quinolone Complex

Kampranis¹,

Maxwell

1998

Journal of Biological Chemistry

109

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Quinolone binding to the gyrase-DNA complex induces a conformational change that results in the blocking of supercoiling. Under these conditions gyrase is still capable of ATP hydrolysis which now proceeds through an alternative pathway involving two different conformations of the enzyme (Kampranis, S. C., and Maxwell, A. (1998) J. Biol. Chem. 269, 22606 -22614). The kinetics of ATP hydrolysis via this pathway have been studied and found to differ from those of the reaction of the drug-free enzyme. The quinolone-characteristic ATPase rate is DNA-dependent and can be induced in the presence of DNA fragments as small as 20 base pairs. By observing the conversion of the ATPase rate to the quinolone characteristic rate, the formation and dissociation of the gyrase-DNA-quinolone complex can be monitored. Comparison of the time dependence of the conversion of the gyrase ATPase with that of DNA cleavage reveals that formation of the gyrase-DNA-quinolone complex does not correspond to the formation of cleaved DNA. Quinolone-induced DNA cleavage proceeds via a mechanism consisting of two cleavage events that is modulated in the presence of a nucleotide cofactor. We demonstrate that quinolone binding and drug-induced DNA cleavage are separate processes constituting two sequential steps in the mechanism of action of quinolones on DNA gyrase.DNA gyrase is the intracellular target of the quinolone group of antibacterial agents. The addition of quinolone drugs to an in vitro reaction containing DNA gyrase, relaxed closed-circular DNA, and ATP leads to the inhibition of supercoiling (2). If this reaction is terminated by the addition of a protein denaturant, like SDS, the DNA is found to be cleaved in both strands with a gyrase A subunit (GyrA) 1 covalently attached to each 5Ј-phosphoryl terminus (3). Quinolones also inhibit the relaxation, catenation, and decatenation reactions of gyrase (2, 4).The molecular details of the interaction of the drugs with the gyrase-DNA complex are unknown. When the binding of the drugs to the enzyme was investigated, insignificant binding of quinolones to the A or B subunit (GyrB) or to the gyrase holoenzyme (A 2 B 2 ) was detected (5-7). Binding of the drugs to the DNA is weak, and more efficient in the case of singlecompared with double-stranded DNA (8,9). Quinolones were found to bind strongly to the complex formed by gyrase and DNA, and ATP appeared to assist this interaction (5). These results led to the proposal that the drugs bind to the singlestranded DNA region that is revealed when gyrase cleaves DNA during turnover (10). This proposal was addressed by site-directed mutagenesis of the gyrase active site. Mutation of tyrosine 122 to serine or phenylalanine abolished supercoiling and quinolone-induced DNA cleavage but the mutants could still bind the drugs equally well as wild-type (11). Clearly DNA cleavage is not required for drug binding. Similar results have also been obtained with topoisomerase IV (12).Using limited proteolysis we found that quinolones stabilize a conformational ...

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Section: Discussionmentioning

confidence: 77%

The DNA Gyrase-Quinolone Complex

Kampranis¹,

Maxwell

1998

Journal of Biological Chemistry

109

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“…The antitumour effects of DOX results from preventing DNA replication and repair leading to cell cycle arrest and apoptosis (Capranico et al, 1990;Gewirtz, 1991;Binaschi et al, 1997). Doxorubicin (DOX)-induced cardiotoxicity was related to the activation of apoptosis in cardiomyocytes and endothelial cells (Wu et al, 2002;Yamanaka et al, 2003;Spallarossa et al, 2004).…”

Section: Discussionmentioning

confidence: 99%

“…Doxorubicin (DOX) induces pleiotropic cytotoxic effects of which DNA intercalation and topoisomerase II inhibition have been proposed to play an important role in its mechanism of action, causing growth arrest and the subsequent activation of apoptosis (Capranico et al, 1990;Gewirtz, 1991;Binaschi et al, 1997). The contribution of DOX-induced reactive oxygen species (ROS) to its antitumour activity is still a matter of debate (Keizer et al, 1991;Shacter et al, 2000;Gouaze et al, 2001;Suresh et al, 2003).…”

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confidence: 99%

Caspase-dependent and -independent suppression of apoptosis by monoHER in Doxorubicin treated cells

et al. 2007

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Doxorubicin (DOX) is an antitumour agent for different types of cancer, but the dose-related cardiotoxicity limits its clinical use. To prevent this side effect we have developed the flavonoid monohydroxyethylrutoside (monoHER), a promising protective agent, which did not interfere with the antitumour activity of DOX. To obtain more insight in the mechanism underlying the selective protective effects of monoHER, we investigated whether monoHER (1 mM) affects DOX-induced apoptosis in neonatal rat cardiac myocytes (NeRCaMs), human endothelial cells (HUVECs) and the ovarian cancer cell lines A2780 and OVCAR-3. DOX-induced cell death was effectively reduced by monoHER in heart, endothelial and A2780 cells. OVCAR-3 cells were highly resistant to DOXinduced apoptosis. Experiments with the caspase-inhibitor zVAD-fmk showed that DOX-induced apoptosis was caspase-dependent in HUVECs and A2780 cells, whereas caspase-independent mechanisms seem to be important in NeRCaMs. MonoHER suppressed DOX-dependent activation of the mitochondrial apoptotic pathway in normal and A2780 cells as illustrated by p53 accumulation and activation of caspase-9 and -3 cleavage. Thus, monoHER acts by suppressing the activation of molecular mechanisms that mediate either caspase-dependent or -independent cell death. In light of the current work and our previous studies, the use of clinically achievable concentrations of monoHER has no influence on the antitumour activity of DOX whereas higher concentrations as used in the present study could influence the antitumour activity of DOX.

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“…Stacking of topoisomerase poisons with the DNA base pairs immediately flanking the topoisomerase cleavage site was initially hypothesized as the mechanism of poisoning of topoisomerase II by several inhibitors of this enzyme (34)(35)(36)(37)(38). The base-stacking hypothesis has been extended to top1 poisons (camptothecins) (6,39).…”

Section: Discussionmentioning

confidence: 99%

Position-specific trapping of topoisomerase I–DNA cleavage complexes by intercalated benzo[ a ]- pyrene diol epoxide adducts at the 6-amino group of adenine

Pommier

Laco

Kohlhagen

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

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DNA topoisomerase I (top1) is the target of potent anticancer agents, including camptothecins and DNA intercalators, which reversibly stabilize (trap) top1 catalytic intermediates (cleavage complexes). The aim of the present study was to define the structural relationship between the site(s) of covalently bound intercalating agents, whose solution conformations in DNA are known, and the site(s) of top1 cleavage. Two diastereomeric pairs of oligonucleotide 22-mers, derived from a sequence used to determine the crystal structure of top1-DNA complexes, were synthesized. One pair contained either a trans-opened 10R-or 10S-benzo[a]pyrene 7,8-diol 9,10-epoxide adduct at the N 6 -amino group of a central 2 -deoxyadenosine residue in the scissile strand, and the other pair contained the same two adducts in the nonscissile strand. These adducts were derived from the (؉)-(7R,8S,9S,10R)-and (؊)-(7S,8R,9R,10S)-7,8-diol 9,10-epoxides in which the benzylic 7-hydroxyl group and the epoxide oxygen are trans. On the basis of analogy with known solution conformations of duplex oligonucleotides containing these adducts, we conclude that top1 cleavage complexes are trapped when the hydrocarbon adduct is intercalated between the base pairs flanking a preexisting top1 cleavage site, or between the base pairs immediately downstream (3 relative to the scissile strand) from this site. We propose a model with the ؉1 base rotated out of the duplex, and in which the intercalated adduct prevents religation of the corresponding nucleotide at the 5 end of the cleaved DNA. These results suggest mechanisms whereby intercalating agents interfere with the normal function of human top1. D NA topoisomerase I (top1) is an essential enzyme in higher eukaryotes (1, 2). It can relax DNA supercoiling and relieve torsional strain during DNA processing, including replication, transcription, and repair. It can also perform intermolecular religation leading to DNA recombinations (3, 4). The catalytic intermediate of top1 is a cleavage complex in which a tyrosine (Tyr-723 for human top1) in the enzyme attacks a DNA phosphodiester and forms a covalent bond to the phosphorus at the 3Ј side of the cleavage site while a 5Ј-hydroxyl is generated at the other side (1, 2). The reaction occurs with short duplex oligodeoxynucleotides (5, 6), as evidenced by recent crystal structures of top1-DNA complexes (7,8).top1 is the cellular target of several anticancer drugs, including the camptothecins, which have recently been approved by the Food and Drug Administration for the treatment of colon and ovarian carcinomas. The anticancer activity of these drugs results from the trapping of top1 cleavage complexes such that the enzyme is reversibly inactivated as it cleaves DNA (9-11). These agents are often referred to as ''top1 poisons.'' They represent an exemplary case of pharmacological interference whereby the drug stabilizes the interactions of two macromolecules (i.e., top1 and DNA) by formation of a ternary complex. Because of the clinical efficacy of camptothecins, no...

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Local sequence requirements for DNA cleavage by mammalian topoisomerase II in the presence of doxorubicin

Cited by 175 publications

References 27 publications

The DNA Gyrase-Quinolone Complex

The DNA Gyrase-Quinolone Complex

Caspase-dependent and -independent suppression of apoptosis by monoHER in Doxorubicin treated cells

Position-specific trapping of topoisomerase I–DNA cleavage complexes by intercalated benzo[ a ]- pyrene diol epoxide adducts at the 6-amino group of adenine

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