Dissecting the Cell-killing Mechanism of the Topoisomerase II-targeting Drug ICRF-193

Oestergaard, Vibe H.; Knudsen, Birgitta R.; Andersen, Anders H.

doi:10.1074/jbc.m402119200

Cited by 19 publications

(21 citation statements)

References 27 publications

(43 reference statements)

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“…In the latter enzyme, the active site tyrosine has been substituted with a serine and the enzyme can neither perform DNA relaxation nor DNA cleavage to a detectable level (29). The extent of stimulation obtained with Y805S is very similar to that obtained with 351i, showing that the DNA cleavage reaction per se does not contribute significantly to the stimulation of topoisomerase II-mediated ATP hydrolysis (Fig.…”

Section: Fig 2 351i Retains Dna Cleavage/ligation Activity a Uppersupporting

confidence: 53%

Hindering the Strand Passage Reaction of Human Topoisomerase IIα without Disturbing DNA Cleavage, ATP Hydrolysis, or the Operation of the N-terminal Clamp

Oestergaard

Giangiacomo²,

Bjergbæk³

et al. 2004

Journal of Biological Chemistry

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DNA topoisomerase II is an essential enzyme that releases a topological strain in DNA by introduction of transient breaks in one DNA helix through which another helix is passed. While changing DNA topology, ATP is required to drive the enzyme through a series of conformational changes dependent on interdomain communication. We have characterized a human topoisomerase II␣ enzyme with a two-amino acid insertion at position 351 in the transducer domain. The mutation specifically abolishes the DNA strand passage event of the enzyme, probably because of a sterical hindrance of T-segment transport. Thus, the enzyme fails to decatenate and relax DNA, even though it is fully capable of ATP hydrolysis, closure of the N-terminal clamp, and DNA cleavage. The cleavage activity is increased, suggesting that the transducer domain has a role in regulating DNA cleavage. Furthermore, the enzyme has retained a tendency to increase DNA cleavage upon nucleotide binding and also responds to DNA with elevated ATP hydrolysis. However, the DNA-mediated increase in ATP hydrolysis is lower than that obtained with the wild-type enzyme but similar to that of a cleavage-deficient topoisomerase II␣ enzyme. Our results strongly suggest that the strand passage event is required for efficient DNA stimulation of topoisomerase II-mediated ATP hydrolysis, whereas the stimulation occurs independent of the DNA cleavage reaction per se. A comparison of the strand passage deficient-enzyme described here and the cleavage-deficient enzyme may have applications in other studies where a clear distinction between strand passage and topoisomerase II-mediated DNA cleavage is desirable.Type II DNA topoisomerases are highly complex enzymes that are able to change the topological conformation of DNA by introducing a transient double strand break in one DNA helix, the G-segment, whereas another intact helix, the T-segment, is transported coordinately through the gated DNA (1). The process depends on ATP, which drives the enzyme through a series of conformational changes (2-5).The homodimeric eukaryotic topoisomerase II enzyme contains two highly conserved catalytic entities, which also share homology with the bacterial DNA topoisomerase II counterpart, DNA gyrase. The ATPase activity is found in the Nterminal region, whereas the DNA cleavage and ligation activities are held by the central region (6). The extreme C termini of the type II topoisomerases are divergent and dispensable for catalytic activity in vitro (7-9).The structural data reveal that the N-terminal region of topoisomerase II forms an ATP-operated clamp, which closes upon ATP binding (10, 11). The dimeric N-terminal clamp contains two domains in each protomer (10). The most Nterminal domain is responsible for dimer interactions during clamp closure and also holds the ATP-binding site. The second domain, called the transducer domain, forms the walls of the clamp and connects it to the enzyme core. Furthermore, communication between the ATP-binding domain and the central domain of the enzyme, respon...

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Section: Fig 2 351i Retains Dna Cleavage/ligation Activity a Uppersupporting

confidence: 53%

Hindering the Strand Passage Reaction of Human Topoisomerase IIα without Disturbing DNA Cleavage, ATP Hydrolysis, or the Operation of the N-terminal Clamp

Oestergaard

Giangiacomo²,

Bjergbæk³

et al. 2004

Journal of Biological Chemistry

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“…Bisdioxopiperazines, including ICRF-187 and ICRF-193, trap eukaryotic topoisomerase II as a closed protein clamp on DNA, which is toxic to cells (Ishida et al, 1995;van Hille and Hill, 1998;Jensen et al, 2000a;Xiao et al, 2003;Oestergaard et al, 2004). This configuration of topoisomerase II on DNA has furthermore been associated with increased levels of DNA strand breaks in some reports (Huang et al, 2001;Hajji et al, 2003).…”

Section: Discussionmentioning

confidence: 99%

A Mouse Model for Studying the Interaction of Bisdioxopiperazines with Topoisomerase IIα in Vivo

Grauslund¹,

Thougaard²,

Füchtbauer³

et al. 2007

Mol Pharmacol

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The bisdioxopiperazines such as (ϩ)-(S)-4,4Ј-propylenedi-2,6-piperazinedione (dexrazoxane; ICRF-187), 1,2-bis(3,5-dioxopiperazin-1-yl)ethane , and 4,4Ј-(1,2-dimethyl-1,2-ethanediyl)bis-2,6-piperazinedione are agents that inhibit eukaryotic topoisomerase II, whereas their ring-opened hydrolysis products are strong iron chelator. The clinically approved analog ICRF-187 is a pharmacological modulator of topoisomerase II poisons such as etoposide in preclinical animal models. ICRF-187 is also used to protect against anthracycline-induced cardiomyopathy and has recently been approved as an antidote for alleviating tissue damage and necrosis after accidental anthracycline extravasation. This dual modality of bisdioxopiperazines, including ICRF-187, raises the question of whether their pharmacological in vivo effects are mediated through interaction with topoisomerase II or via their intracellular iron chelating activity. In an attempt to distinguish between these possibilities, we here present a transgenic mouse model aimed at identifying the contribution of topoisomerase II␣ to the effects of bisdioxopiperazines. A tyrosine 165 to serine mutation (Y165S) in topoisomerase II␣, demonstrated previously to render the human ortholog of this enzyme highly resistant toward bisdioxopiperazines, was introduced at the TOP2A locus in mouse embryonic stem cells by targeted homologous recombination. These cells were used for the generation of transgenic TOP2A Y165S/ϩ mice, which were demonstrated to be resistant toward the general toxicity of both . Hematological measurements indicate that this is most likely caused by a decreased ability of these agents to induce myelosuppression in TOP2A Y165S/ϩ mice, highlighting the role of topoisomerase II␣ in this process. The biological and pharmacological implications of these findings are discussed, and areas for further investigations are proposed.The nuclear enzyme topoisomerase II is found in all living organisms and possesses the essential activity of cleaving and rejoining DNA by forming a transient protein concealed double-strand break, through which an intact DNA doublestrand helix is transported by a gating mechanism requiring ATP hydrolysis Baird et al., 2001). This activity assists in a number of DNA metabolic processes such as DNA replication, transcription, chromosome condensation and decondensation, and chromosome segregation (Wang, 2002). Higher vertebrates have two isoforms of this enzyme, ␣ and ␤. The topoisomerase II␣ isoform is nuclear, and its expression is highly cell cycle-dependent, peaking at G 2 M , whereas topoisomerase II␤ is located both in the nucleus and cytoplasm and displays a rather constant level during the cell cycle (Austin and Marsh, 1998). Topoisomerase II␣ is essential for cell proliferation (Akimitsu et al., 2003), whereas topoisomerase II ␤ knockout mice die shortly after birth because of developmental defects in the central nervous system (CNS) (Yang et al., 2000).The bisdioxopiperazines such as Morris et al., 2000). This closed-clamp ...

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“…These results point towards bisdioxopiperazines poisoning DNA topoisomerase II in cells by a mechanism different from that of the classical topoisomerase II poisons such as etoposide and m-AMSA. In a recent paper, it was directly demonstrated that m-AMSA-induced dominant cytotoxicity only required the DNA cleavage activity of topoisomerase II, while dominant cytotoxicity towards ICRF-193 depended strictly on the DNA strand passage reaction of the enzyme[17]. …”

Section: Introductionmentioning

confidence: 99%

Characterisation of cytotoxicity and DNA damage induced by the topoisomerase II-directed bisdioxopiperazine anti-cancer agent ICRF-187 (dexrazoxane) in yeast and mammalian cells

Jensen¹,

Dejligbjerg²,

Hansen

et al. 2004

BMC Pharmacol

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Background: Bisdioxopiperazine anti-cancer agents are inhibitors of eukaryotic DNA topoisomerase II, sequestering this protein as a non-covalent protein clamp on DNA. It has been suggested that such complexes on DNA represents a novel form of DNA damage to cells. In this report, we characterise the cytotoxicity and DNA damage induced by the bisdioxopiperazine ICRF-187 by a combination of genetic and molecular approaches. In addition, the well-established topoisomerase II poison m-AMSA is used for comparison.

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Dissecting the Cell-killing Mechanism of the Topoisomerase II-targeting Drug ICRF-193

Cited by 19 publications

References 27 publications

Hindering the Strand Passage Reaction of Human Topoisomerase IIα without Disturbing DNA Cleavage, ATP Hydrolysis, or the Operation of the N-terminal Clamp

Hindering the Strand Passage Reaction of Human Topoisomerase IIα without Disturbing DNA Cleavage, ATP Hydrolysis, or the Operation of the N-terminal Clamp

A Mouse Model for Studying the Interaction of Bisdioxopiperazines with Topoisomerase IIα in Vivo

Characterisation of cytotoxicity and DNA damage induced by the topoisomerase II-directed bisdioxopiperazine anti-cancer agent ICRF-187 (dexrazoxane) in yeast and mammalian cells

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