Abstract
The metabolite (−)-lomaiviticin A, which contains two diazotetrahydrobenzo[b]fluorene (diazofluorene) functional groups, inhibits the growth of cultured human cancer cells at nanomolar–picomolar concentrations; however, the mechanism responsible for the potent cytotoxicity of this natural product is not known. Here we report that (−)-lomaiviticin A nicks and cleaves plasmid DNA by a pathway that is independent of reactive oxygen species and iron, and that the potent cytotoxicity of (−)-lomaiviticin A arises from the induction of DNA double-strand breaks (dsbs). In a plasmid cleavage assay, the ratio of single-strand breaks (ssbs) to dsbs is 5.3 ± 0.6:1. Labelling studies suggest that this cleavage occurs via a radical pathway. The structurally related isolates (−)-lomaiviticin C and (−)-kinamycin C, which contain one diazofluorene, are demonstrated to be much less effective DNA cleavage agents, thereby providing an explanation for the enhanced cytotoxicity of (−)-lomaiviticin A compared to that of other members of this family.
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Acknowledgements
The authors dedicate this paper to the memory of their colleague Donald M. Crothers. Financial support from the National Institute of General Medical Sciences (R01GM090000, S.B.H.), the National Institute of Environmental Health Sciences (R01ES005775, P.M.G.), the National Cancer Institute (R01CA168733, P.M.G.), the National Science Foundation (Graduate Research Fellowship to C.M.W.), the Searle Scholars Program (S.B.H.) and Yale University (S.B.H.) is gratefully acknowledged. S.B.H. acknowledges early-stage investigator awards from the David and Lucile Packard Foundation, the Alfred P. Sloan Foundation, the Camille and Henry Dreyfus Foundation, and the Research Corporation for Science Advancement. We thank T. Dymarz, J. Shen, D. Spiegel and T. Wang for assistance.
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L.C.C. and C.M.W. designed and performed the plasmid cleavage, immunofluorescence, comet and flow cytometry experiments. Z.L. and C.M.W. performed the in vitro reactivity studies. P.M.G. and D.A.H. designed, performed and analysed the clonogenic survival assays, pATM/pChk2/pATR/pChk1 western blots and comet assay employing BRCA2-deficient cells. S.B.H. conceived and designed the study, analysed the data and composed the manuscript.
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Colis, L., Woo, C., Hegan, D. et al. The cytotoxicity of (−)-lomaiviticin A arises from induction of double-strand breaks in DNA. Nature Chem 6, 504–510 (2014). https://doi.org/10.1038/nchem.1944
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DOI: https://doi.org/10.1038/nchem.1944
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