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Effects of novel inhibitors of poly(ADP-ribose) polymerase-1 and the DNA-dependent protein kinase on enzyme activities and DNA repair

Oncogene volume 23, pages 7322–7329 (2004)Cite this article

Abstract

DNA-dependent protein kinase (DNA-PK) and poly (ADP-ribose) polymerase-1 (PARP-1) participate in nonhomologous end joining and base excision repair, respectively, and are key determinants of radio- and chemo-resistance. Both PARP-1 and DNA-PK have been identified as therapeutic targets for anticancer drug development. Here we investigate the effects of specific inhibitors on enzyme activities and DNA double-strand break (DSB) repair. The enzyme activities were investigated using purified enzymes and in permeabilized cells. Inhibition, or loss of activity, was compared using potent inhibitors of DNA-PK (NU7026) and PARP-1 (AG14361), and cell lines proficient or deficient for DNA-PK or PARP-1. Inactive DNA-PK suppressed the activity of PARP-1 and vice versa. This was not the consequence of simple substrate competition, since DNA ends were provided in excess. The inhibitory effect of DNA-PK on PARP activity was confirmed in permeabilized cells. Both inhibitors prevented ionizing radiation-induced DSB repair, but only AG14361 prevented single-strand break repair. An increase in DSB levels caused by inhibition of PARP-1 was shown to be caused by a decrease in DSB repair, and not by the formation of additional DSBs. These data point to combined inhibition of PARP-1 and DNA-PK as a powerful strategy for tumor radiosensitization.

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Acknowledgements

This work was supported by funds from the Cancer Research UK. AG14361 was kindly provided by Pfizer GRD, San Diego, California.

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Authors and Affiliations

  1. Northern Institute for Cancer Research, University of Newcastle, Newcastle upon Tyne, NE2 4HH, UK

    Stephany J Veuger, Nicola J Curtin & Barbara W Durkacz

  2. KuDOS Pharmaceuticals, Cambridge Science Park, Cambridge, CB4 4WG, UK

    Graeme CM Smith

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  1. Stephany J Veuger
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  4. Barbara W Durkacz
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Correspondence to Barbara W Durkacz.

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Veuger, S., Curtin, N., Smith, G. et al. Effects of novel inhibitors of poly(ADP-ribose) polymerase-1 and the DNA-dependent protein kinase on enzyme activities and DNA repair. Oncogene 23, 7322–7329 (2004). https://doi.org/10.1038/sj.onc.1207984

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