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Cleaving the oxidative repair protein Ape1 enhances cell death mediated by granzyme A

Nature Immunology volume 4, pages 145–153 (2003)Cite this article

Abstract

The cytolytic T lymphocyte protease granzyme A (GzmA) initiates a caspase-independent cell death pathway. Here we report that the rate-limiting enzyme of DNA base excision repair, apurinic endonuclease-1 (Ape1), which is also known as redox factor-1 (Ref-1), binds to GzmA and is contained in the SET complex, a macromolecular complex of 270–420 kDa that is associated with the endoplasmic reticulum and is targeted by GzmA during cell-mediated death. GzmA cleaves Ape1 after Lys31 and destroys its known oxidative repair functions. In so doing, GzmA may block cellular repair and force apoptosis. In support of this, cells with silenced Ape1 expression are more sensitive, whereas cells overexpressing noncleavable Ape1 are more resistant, to GzmA-mediated death.

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Figure 1: Ape1 co-elutes with the SET complex.
Figure 2: Ape1 interacts and colocalizes with pp32 and SET.
Figure 3: Ape1 is a substrate of GzmA but not GzmB.
Figure 4: GzmA and PFP treatment of HeLa cells causes degradation of Ape1.
Figure 5: GzmA disrupts the AP endonuclease and redox activities of Ape1.
Figure 6: Silencing of Ape1 expression enhances GzmA-mediated cytolysis and DNA nicking.
Figure 7: Overexpressing noncleavable Ape1 reduces GzmA-induced cell death.

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Acknowledgements

We thank M. Russo for technical support; B. Demple, D. Oh, X.-F. Yang, L. Shi and K. Kaznatcheev for suggestions; and the National Center for Cell Culture for preparing K562 cell lysates to purify the SET complex. This work was supported by a grant from the National Institutes of Health (NIH) to J.L. and the NIH-supported Dana Farber Cancer Institute, Beth Israel Deaconess Medical Center and Children's Hospital Center for AIDS Research.

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Author notes

  1. Akira Yoshida

    Present address: Department of Internal Medicine, Fukui Medical University, Matsuoka, 910-1193, Japan

Authors and Affiliations

  1. Center for Blood Research, Harvard Medical School, Boston, 02115, MA, USA

    Zusen Fan, Paul J. Beresford, Dong Zhang, Zhan Xu & Judy Lieberman

  2. Department of Pediatrics, Harvard Medical School, Boston, 02115, MA, USA

    Zusen Fan, Paul J. Beresford, Dong Zhang & Judy Lieberman

  3. Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, 02139, MA, USA

    Carl D. Novina

  4. Laboratory of Molecular Pharmacology, National Cancer Institute, Bethesda, 20892, MD, USA

    Akira Yoshida & Yves Pommier

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Fan, Z., Beresford, P., Zhang, D. et al. Cleaving the oxidative repair protein Ape1 enhances cell death mediated by granzyme A. Nat Immunol 4, 145–153 (2003). https://doi.org/10.1038/ni885

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