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The lyase activity of the DNA repair protein β-polymerase protects from DNA-damage-induced cytotoxicity

Nature volume 405, pages 807–810 (2000)Cite this article

Abstract

Small DNA lesions such as oxidized or alkylated bases are repaired by the base excision repair (BER) pathway1. BER includes removal of the damaged base by a lesion-specific DNA glycosylase, strand scission by apurinic/apyrimidinic endonuclease, DNA resynthesis and ligation2. BER may be further subdivided into DNA β-polymerase (β-pol)-dependent single-nucleotide repair and β-pol-dependent or -independent long patch repair subpathways3,4,5,6. Two important enzymatic steps in mammalian single-nucleotide BER are contributed by β-pol: DNA resynthesis of the repair patch and lyase removal of 5′-deoxyribose phosphate (dRP)2. Fibroblasts from β-pol null mice are hypersensitive to monofunctional DNA-methylating agents, resulting in increases in chromosomal damage, apoptosis and necrotic cell death3,7. Here we show that only the dRP lyase activity of β-pol is required to reverse methylating agent hypersensitivity in β-pol null cells. These results indicate that removal of the dRP group is a pivotal step in BER in vivo. Persistence of the dRP moiety in DNA results in the hypersensitivity phenotype of β-pol null cells and may signal downstream events such as apoptosis and necrotic cell death.

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Figure 1: Sensitivity of fibroblast cell lines to MMS.
Figure 2: DNA synthesis activity of β-pol is not required to reverse the MMS hypersensitivity of β-pol null cells.
Figure 3: RP lyase activity of β-pol is sufficient to reverse the MMS hypersensitivity of β-pol null cells.
Figure 4: RP lyase activity in whole-cell extracts from stably transfected β-pol null cell lines.

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Acknowledgements

We thank W. A. Beard and K. R. Tindall for critical reading of the manuscript.

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  1. Laboratory of Structural Biology, National Institute of Environmental Health Sciences, PO Box 12233, Research Triangle Park, 27709-2233, North Carolina, USA

    Robert W. Sobol, Rajendra Prasad, Andrea Evenski, Audrey Baker, Xiao-Ping Yang, Julie K. Horton & Samuel H. Wilson

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  1. Robert W. Sobol
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Correspondence to Samuel H. Wilson.

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Sobol, R., Prasad, R., Evenski, A. et al. The lyase activity of the DNA repair protein β-polymerase protects from DNA-damage-induced cytotoxicity. Nature 405, 807–810 (2000). https://doi.org/10.1038/35015598

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