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Basis for recognition of cisplatin-modified DNA by high-mobility-group proteins

Nature volume 399, pages 708–712 (1999)Cite this article

Abstract

The anticancer activity of cis -diamminedichloroplatinum(II) (cisplatin) arises from its ability to damage DNA, with the major adducts formed being intrastrand d(GpG) and d(ApG) crosslinks1. These crosslinks bend and unwind the duplex, and the altered structure attracts high-mobility-group domain (HMG) and other proteins2. This binding of HMG-domain proteins to cisplatin-modified DNA has been postulated to mediate the antitumour properties of the drug3,4. Many HMG-domain proteins recognize altered DNA structures such as four-way junctions and cisplatin-modified DNA5, but until now the molecular basis for this recognition was unknown. Here we describe mutagenesis, hydroxyl-radical footprinting and X-ray studies that elucidate the structure of a 1:1 cisplatin-modified DNA/HMG-domain complex. Domain A of the structure-specific HMG-domain protein HMG1 binds to the widened minor groove of a 16-base-pair DNA duplex containing a site-specific cis -[Pt(NH3)2{d(GpG)-N7(1),-N7(2)}] adduct. The DNA is strongly kinked at a hydrophobic notch created at the platinum–DNA crosslink and protein binding extends exclusively to the 3′ side of the platinated strand. A phenylalanine residue at position 37 intercalates into a hydrophobic notch created at the platinum crosslinked d(GpG) site and binding of the domain is dramatically reduced in a mutant in which alanine is substituted for phenylalanine at this position.

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Figure 1: Primary structure and features of the complex between the non-sequence-specific domain A of HMG1 and cisplatin-modified DNA.
Figure 2: Structure of the HMG1 domain A complex with cisplatin-modified DNA.
Figure 3: The platinum–DNA crosslink and protein–DNA interactions.

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Acknowledgements

This work was supported by a grant from the National Cancer Institute (to S.J.L.), by the Howard Hughes Medical Institute (C.O.P.), by equipment purchased with support from the PEW Charitable Trusts, and by predoctoral fellowships from the Fonds der Chemischen Industrie, Germany (U.-M.O.), and the Howard Hughes Medical Institute (Q.H.). We thank S. U. Dunham for discussion, and J. S. Wishnok for mass spectrometric analysis.

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  1. Mark A. Rould

    Present address: Department of Molecular Physiology and Biophysics, University of Vermont College of Medicine, Burlington, Vermont, 05405, USA

Authors and Affiliations

  1. Department of Chemistry, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts, USA

    Uta-Maria Ohndorf, Qing He & Stephen J. Lippard

  2. Department of Biology and Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts, USA

    Mark A. Rould & Carl O. Pabo

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Correspondence to Stephen J. Lippard.

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Ohndorf, UM., Rould, M., He, Q. et al. Basis for recognition of cisplatin-modified DNA by high-mobility-group proteins. Nature 399, 708–712 (1999). https://doi.org/10.1038/21460

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