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Structural basis for cisplatin DNA damage tolerance by human polymerase η during cancer chemotherapy

Nature Structural & Molecular Biology volume 19, pages 628–632 (2012)Cite this article

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Abstract

A major clinical problem in the use of cisplatin to treat cancers is tumor resistance. DNA polymerase η (Pol-η) is a crucial polymerase that allows cancer cells to cope with the cisplatin–DNA adducts that are formed during chemotherapy. We present here a structure of human Pol-η inserting deoxycytidine triphosphate (dCTP) opposite a cisplatin intrastrand cross-link (PtGpG). We show that the specificity of human Pol-η for PtGpG derives from an active site that is open to permit Watson-Crick geometry of the nascent PtGpG-dCTP base pair and to accommodate the lesion without steric hindrance. This specificity is augmented by the residues Gln38 and Ser62, which interact with PtGpG, and Arg61, which interacts with the incoming dCTP. Collectively, the structure provides a basis for understanding how Pol-η in human cells can tolerate the DNA damage caused by cisplatin chemotherapy and offers a framework for the design of inhibitors in cancer therapy.

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Figure 1: Human Pol-η–DNA ternary complexes.
Figure 2: Comparison of the human Pol-η–PtGpG ternary complex with the human Pol-η–thymine–thymine dimer and human Pol-κ–DNA complexes.
Figure 3: Close-up views of the active site regions.

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Acknowledgements

We thank the staff at Brookhaven National Laboratory (beamline X25) for facilitating X-ray data collection. We also thank G. Gerona-Navarro for assistance with HPLC. This work was supported by grants ES017767 and ES012411 from the US National Institutes of Health. A.U. was supported in part by US National Institutes of Health training grant T32 GM62754.

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Authors and Affiliations

  1. Department of Structural and Chemical Biology, Mount Sinai School of Medicine, New York, New York, USA

    Ajay Ummat, Olga Rechkoblit, Rinku Jain, Timothy D Silverstein, Angeliki Buku, Samer Lone & Aneel K Aggarwal

  2. Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, Texas, USA

    Jayati Roy Choudhury, Robert E Johnson, Louise Prakash & Satya Prakash

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  1. Ajay Ummat
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Contributions

A.K.A. and A.U. designed the experiments. A.U. determined the crystal structure. O.R. and A.U. prepared the cisplatin adduct. R.J. assisted in the crystallographic analysis. T.D.S. and S.L. assisted in DNA preparation. A.B. assisted in protein purification. J.R.C. and R.E.J. conducted the biochemical experiments. L.P. and S.P. guided the biochemical studies. A.K.A. wrote the manuscript with contributions from all of the authors.

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Correspondence to Aneel K Aggarwal.

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The authors declare no competing financial interests.

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Supplementary Figures 1–3 and Supplementary Table 1 (PDF 2332 kb)

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Ummat, A., Rechkoblit, O., Jain, R. et al. Structural basis for cisplatin DNA damage tolerance by human polymerase η during cancer chemotherapy. Nat Struct Mol Biol 19, 628–632 (2012). https://doi.org/10.1038/nsmb.2295

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