Anti-tumour compounds illudin S and Irofulven induce DNA lesions ignored by global repair and exclusively processed by transcription- and replication-coupled repair pathways

Jaspers, Nicolaas G. J.; Raams, Anja; Kelner, Michael J.; Ng, Jessica M.Y.; Yamashita, Yukiko; Takeda, Shunichi; McMorris, Trevor C.; Hoeijmakers, Jan H.J.

doi:10.1016/s1568-7864(02)00166-0

Cited by 147 publications

(164 citation statements)

References 51 publications

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“…Cds1 activation triggers Rad60 phosphorylation and nuclear delocalization regulating recombination events at stalled replication forks (95). While it is yet to be determined whether CHK2 activation might also play a role in repairing irofulven-elicited DNA lesions, it has been reported that irofulven-induced DNA damage is repaired by transcription-coupled nucleotide excision repair (TC-NER) (21). Therefore, it can be speculated that the CHK2 activation by irofulven might be the result of stalled replication machinery.…”

Section: Discussionmentioning

confidence: 99%

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ATM-dependent CHK2 Activation Induced by Anticancer Agent, Irofulven

Wang¹,

Wiltshire

Wang³

et al. 2004

Journal of Biological Chemistry

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Irofulven (6-hydroxymethylacylfulvene, HMAF, MGI 114) is one of a new class of anticancer agents that are semisynthetic derivatives of the mushroom toxin illudin S. Preclinical studies and clinical trials have demonstrated that irofulven is effective against several tumor types. Mechanisms of action studies indicate that irofulven induces DNA damage, MAPK activation, and apoptosis. In this study we found that in ovarian cancer cells, CHK2 kinase is activated by irofulven while CHK1 kinase is not activated even when treated at higher concentrations of the drug. By using GM00847 human fibroblast expressing tetracycline-controlled, FLAG-tagged kinase-dead ATR (ATR.kd), it was demonstrated that ATR kinase does not play a major role in irofulveninduced CHK2 activation. Results from human fibroblasts proficient or deficient in ATM function (GM00637 and GM05849) indicated that CHK2 activation by irofulven is mediated by the upstream ATM kinase. Phosphorylation of ATM on Ser 1981 , which is critical for kinase activation, was observed in ovarian cancer cell lines treated with irofulven. RNA interference results confirmed that CHK2 activation was inhibited after introducing siRNA for ATM. Finally, experiments done with human colon cancer cell line HCT116 and its isogenic CHK2 knockout derivative; and experiments done by expressing kinase-dead CHK2 in an ovarian cancer cell line demonstrated that CHK2 activation contributes to irofulven-induced S phase arrest. In addition, it was shown that NBS1, SMC1, and p53 were phosphorylated in an ATM-dependent manner, and p53 phosphorylation on serine 20 is dependent on CHK2 after irofulven treatment. In summary, we found that the anticancer agent, irofulven, activates the ATM-CHK2 DNA damage-signaling pathway, and CHK2 activation contributes to S phase cell cycle arrest induced by irofulven.Irofulven 1 (6-hydroxymethylacylfulvene, HMAF, MGI 114) is one of a new class of anticancer agents that are analogs of mushroom-derived illudin toxins. Preclinical studies and clinical trials have demonstrated that irofulven is effective against several tumor types (1-17). Studies of mechanisms of irofulven action suggest that it induces DNA damage, MAP kinase activation and apoptosis (18 -20). It is also suggested that irofulven-elicited DNA lesions are mainly repaired by transcriptioncoupled nucleotide excision repair (TC-NER) (21).In response to DNA damage, the cell evokes signal transduction pathways to arrest at G 1 /S, S, or G 2 /M checkpoints, allowing time to deal with the insult (22, 23). It has been well documented that DNA damage activates ATM (ataxia telangiectasia-mutated), and/or ATR (ATM-RAD3-related) kinases, two apical protein kinases of the DNA damage response pathways. ATM and ATR phosphorylate downstream effector kinases, CHK1 and CHK2. It is generally believed that ATM is the kinase mainly responding to ionizing radiation (IR)-induced DNA double strand breaks, while ATR responds to the formation of DNA adducts and stalled replication induced by UV, genotoxic drugs, and radia...

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Section: Discussionmentioning

confidence: 99%

“…Studies of mechanisms of irofulven action suggest that it induces DNA damage, MAP kinase activation and apoptosis (18 -20). It is also suggested that irofulven-elicited DNA lesions are mainly repaired by transcriptioncoupled nucleotide excision repair (TC-NER) (21).…”

mentioning

confidence: 99%

ATM-dependent CHK2 Activation Induced by Anticancer Agent, Irofulven

Wang¹,

Wiltshire

Wang³

et al. 2004

Journal of Biological Chemistry

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“…We (Koeppel et al, 2004) and others (Kelner et al, 1994;Jaspers et al, 2002) have reported that irofulven (6-hydroxymethylacylfulvene), a monofunctional covalent DNA binder (Woynarowski et al, 1997), and structurally related compounds induce DNA adducts that are exclusively recognized by TCR, but not by GGR. Here, we report that exposure of mammalian cells to irofulven is associated with potent and specific inhibition of nucleoplasmic RNA synthesis, an important decrease of Pol II mobility and a time-dependent loss of Pol II LS linked to its polyubiquitylation and proteasome-mediated degradation.…”

Section: Introductionmentioning

confidence: 89%

Influence of irofulven, a transcription-coupled repair-specific antitumor agent, on RNA polymerase activity, stability and dynamics in living mammalian cells

Escargueil

Poindessous

Soares

et al. 2008

Journal of Cell Science

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Transcription-coupled repair (TCR) plays a key role in the repair of DNA lesions induced by bulky adducts and is initiated when the elongating RNA polymerase II (Pol II) stalls at DNA lesions. This is accompanied by alterations in Pol II activity and stability. We have previously shown that the monofunctional adducts formed by irofulven (6-hydroxymethylacylfulvene) are exclusively recognized by TCR, without involvement of global genome repair (GGR), making irofulven a unique tool to characterize TCR-associated processes in vivo. Here, we characterize the influence of irofulven on Pol II activity, stability and mobility in living mammalian cells. Our results demonstrate that irofulven induces specific inhibition of nucleoplasmic RNA synthesis, an important decrease of Pol II mobility, coupled to the accumulation of initiating polymerase and a time-dependent loss of the engaged enzyme, associated with its polyubiquitylation. Both proteasome-mediated degradation of the stalled polymerase and new protein synthesis are necessary to allow Pol II recycling into preinitiating complexes. Together, our findings provide novel insights into the subsequent fate of the stalled RNA polymerase II and demonstrate the essential role of the recycling process for transcriptional reinitiation and viability of mammalian cells.

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“…This group includes ecteinascidin-743 (ET-743, Yondelis, trabectedin), irofulven, hedamycin, and the acronycine derivative S23906. Mechanistic studies have revealed several differences in the way these compounds react with DNA-metabolizing enzymes as well as with the repair machinery (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11), which could, at least in part, explain their different activity spectra. A better understanding of the factors controlling the induction and removal of DNA damage induced by these compounds should not only facilitate their clinical application, but may also contribute to our general understanding of the structural and biological features governing adduct processing in mammalian cells.…”

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confidence: 99%

Replication and homologous recombination repair regulate DNA double-strand break formation by the antitumor alkylator ecteinascidin 743

Soares

Escargueil

Poindessous

et al. 2007

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

144

111

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Adducts induced by the antitumor alkylator ecteinascidin 743 (ET-743, Yondelis, trabectedin) represent a unique challenge to the DNA repair machinery because no pathway examined to date is able to remove the ET adducts, whereas cells deficient in nucleotide excision repair show increased resistance. We here describe the processing of the initial ET adducts into cytotoxic lesions and characterize the influence of cellular repair pathways on this process. Our findings show that exposure of proliferating mammalian cells to pharmacologically relevant concentrations of ET-743 is accompanied by rapid formation of DNA double-strand breaks (DSBs), as shown by the neutral comet assay and induction of focalized phosphorylated H2AX. The ET adducts are stable and can be converted into DSBs hours after the drug has been removed. Loss of homologous recombination repair has no influence on the initial levels of DSBs but is associated with the persistence of unrepaired DSBs after ET-743 is removed, resulting in extensive chromosomal abnormalities and pronounced sensitivity to the drug. In comparison, loss of nonhomologous end-joining had only modest effect on the sensitivity. The identification of DSB formation as a key step in the processing of ET-743 lesions represents a novel mechanism of action for the drug that is in agreement with its unusual potency. Because loss of repair proteins is common in human tumors, expression levels of selected repair factors may be useful in identifying patients particularly likely to benefit, or not, from treatment with ET-743.cancer therapy ͉ natural products ͉ lesion processing ͉ DNA repair ͉ response prediction

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Anti-tumour compounds illudin S and Irofulven induce DNA lesions ignored by global repair and exclusively processed by transcription- and replication-coupled repair pathways

Cited by 147 publications

References 51 publications

ATM-dependent CHK2 Activation Induced by Anticancer Agent, Irofulven

ATM-dependent CHK2 Activation Induced by Anticancer Agent, Irofulven

Influence of irofulven, a transcription-coupled repair-specific antitumor agent, on RNA polymerase activity, stability and dynamics in living mammalian cells

Replication and homologous recombination repair regulate DNA double-strand break formation by the antitumor alkylator ecteinascidin 743

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