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The role of Cdk5-mediated apurinic/apyrimidinic endonuclease 1 phosphorylation in neuronal death

Nature Cell Biology volume 12, pages 563–571 (2010)Cite this article

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Abstract

Accumulating evidence suggests that deregulated cyclin-dependent kinase 5 (Cdk5) plays a critical part in neuronal death. However, the pathogenic targets of Cdk5 are not fully defined. Here we demonstrate that the Cdk5 activator p35 interacts directly with apurinic/apyrimidinic endonuclease 1 (Ape1), a protein crucial for base excision repair (BER) following DNA damage. Cdk5 complexes phosphorylate Ape1 at Thr 232 and thereby reduces its apurinic/apyrimidinic (AP) endonuclease activity. Ape1 phosphorylation is dependent on Cdk5 in in vitro and in vivo. The reduced endonuclease activity of phosphorylated Ape1 results in accumulation of DNA damage and contributes to neuronal death. Overexpression of Ape1WT and Ape1T232A, but not the phosphorylation mimic Ape1T232E, protects neurons against MPP+/MPTP. Loss of Ape1 sensitizes neurons to death. Importantly, increased phosphorylated Ape1 was also observed in post-mortem brain tissue from patients with Parkinson's and Alzheimer's diseases, suggesting a potential link between Ape1 phosphorylation and the pathogenesis of neurodegenerative diseases.

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Figure 1: Identification of interaction between Cdk5/p35 and Ape1 in vitro.
Figure 2: The effect of Cdk5-mediated phosphorylation of Ape1 and its mutants on AP endonuclease activity in vitro.
Figure 3: Induction of Ape1 phosphorylation at Thr 232 in MPP+-treated neurons.
Figure 4: The role of Ape1 phosphorylation at Thr 232 in MPP+-treated neurons.
Figure 5: The role of Ape1 phosphorylation at Thr 232 in MPTP–injected mice and mediated by Cdk5.
Figure 6: Ape1 phosphorylation at Thr 232 is increased in human Parkinson's disease and Alzheimer's disease.

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Acknowledgements

This work was partially supported by Parkinson Society Canada (E.H., D.Q.); by the Heart and Stroke Foundation of Ontario (M.W.C.R); and by funds from the Canadian Institutes of Health Research, Parkinson Society Canada, Parkinson's Disease Foundation, Parkinson Research Consortium, Heart and Stroke Foundation of Ontario, Brain Repair Program-Neuroscience Canada, and World Class University Program, National Research Foundation, Ministry of Education, Science & Technology, South Korea, Grant (R31-2008-000-20009-0) (D.S.P.). D.S.P. is a Heart and Stroke Foundation of Ontario career investigator.

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  1. En Huang and Dianbo Qu: These authors contributed equally to this work.

Authors and Affiliations

  1. Cellular Molecular Medicine, Ottawa Hospital Research Institute, University of Ottawa, 451 Smyth Road, Ottawa, K1H 8M5, Ontario, Canada

    En Huang, Dianbo Qu, Yi Zhang, Katerina Venderova, M. Emdadul Haque, Maxime W.C. Rousseaux, Ruth S. Slack & David S. Park

  2. Departments of Pathology and Laboratory Medicine and of Biochemistry, University of Ottawa, Cancer Research Group, OHRI. Ottawa Hospital, Civic Campus, Ottawa, K1Y 4E9, Ontario, Canada

    John M. Woulfe

  3. Department of Cogno-Mechatronics Engineering, Pusan National University, Geumjeong GU, Busan, 609 735, South Korea

    David S. Park

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Contributions

E.H., Q.D., R.S.S., D.S.P. designed the studies; E.H. and D.Q. performed most of the experimental work; Y.Z., K.V., M. E. H. and M.W.C.R. provided technical support;. J. M. W. provided post-mortem Parkinson's disease and Alzheimer's disease samples and analyses; E.H., Q.D. and D.S.P. analysed data and wrote the manuscript; D.S.P supervised the project.

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Correspondence to David S. Park.

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Huang, E., Qu, D., Zhang, Y. et al. The role of Cdk5-mediated apurinic/apyrimidinic endonuclease 1 phosphorylation in neuronal death. Nat Cell Biol 12, 563–571 (2010). https://doi.org/10.1038/ncb2058

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