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Mechanism of transcriptional stalling at cisplatin-damaged DNA

Nature Structural & Molecular Biology volume 14, pages 11271133 (2007) | Download Citation

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Abstract

The anticancer drug cisplatin forms 1,2-d(GpG) DNA intrastrand cross-links (cisplatin lesions) that stall RNA polymerase II (Pol II) and trigger transcription-coupled DNA repair. Here we present a structure-function analysis of Pol II stalling at a cisplatin lesion in the DNA template. Pol II stalling results from a translocation barrier that prevents delivery of the lesion to the active site. AMP misincorporation occurs at the barrier and also at an abasic site, suggesting that it arises from nontemplated synthesis according to an 'A-rule' known for DNA polymerases. Pol II can bypass a cisplatin lesion that is artificially placed beyond the translocation barrier, even in the presence of a G·A mismatch. Thus, the barrier prevents transcriptional mutagenesis. The stalling mechanism differs from that of Pol II stalling at a photolesion, which involves delivery of the lesion to the active site and lesion-templated misincorporation that blocks transcription.

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Acknowledgements

We thank members of the Cramer laboratory for help. P.C. and T.C. were supported by the Deutsche Forschungsgemeinschaft, the Sonderforschungsbereich SFB646 and the Fonds der chemischen Industrie. P.C. was supported by the EU grant 3D repertoire, contract no. LSHG-CT-2005-512028. G.E.D. and F.B. were supported by the Elite Netzwerk Bayern. A.A. was supported by the Marie Curie training and mobility network CLUSTOX DNA.

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Affiliations

  1. Center for Integrated Protein Science CIPSM, Department of Chemistry and Biochemistry, Ludwig-Maximilians-Universität München, Feodor-Lynen-Strasse 25, 81377 Munich, Germany.

    • Gerke E Damsma
    • , Aaron Alt
    • , Florian Brueckner
    • , Thomas Carell
    •  & Patrick Cramer
  2. Gene Center Munich, Ludwig-Maximilians-Universität München, Feodor-Lynen-Strasse 25, 81377 Munich, Germany.

    • Gerke E Damsma
    • , Florian Brueckner
    •  & Patrick Cramer

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Contributions

G.E.D. performed and analyzed biochemical and crystallographic experiments. A.A. synthesized cisplatin-containing DNA strands and performed MALDI experiments. F.B. assisted with experiments and crystallography. T.C. supervised the project. P.C. supervised the project and wrote the manuscript.

Corresponding authors

Correspondence to Thomas Carell or Patrick Cramer.

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https://doi.org/10.1038/nsmb1314

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