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Oxidant-induced apoptosis is mediated by oxidation of the actin-regulatory protein cofilin

Nature Cell Biology volume 11, pages 1241–1246 (2009)Cite this article

A Corrigendum to this article was published on 01 November 2009

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Abstract

Physiological oxidants that are generated by activated phagocytes comprise the main source of oxidative stress during inflammation1,2. Oxidants such as taurine chloramine (TnCl) and hydrogen peroxide (H2O2) can damage proteins and induce apoptosis, but the role of specific protein oxidation in this process has not been defined. We found that the actin-binding protein cofilin is a key target of oxidation. When oxidation of this single regulatory protein is prevented, oxidant-induced apoptosis is inhibited. Oxidation of cofilin causes it to lose its affinity for actin and to translocate to the mitochondria, where it induces swelling and cytochrome c release by mediating opening of the permeability transition pore (PTP). This occurs independently of Bax activation and requires both oxidation of cofilin Cys residues and dephosphorylation at Ser 3. Knockdown of endogenous cofilin using targeted siRNA inhibits oxidant-induced apoptosis, which is restored by re-expression of wild-type cofilin but not by cofilin containing Cys to Ala mutations. Exposure of cofilin to TnCl results in intramolecular disulphide bonding and oxidation of Met residues to Met sulphoxide, but only Cys oxidation causes cofilin to induce mitochondrial damage.

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Figure 1: TnCl causes dissociation of actin/cofilin complexes and translocation of cofilin to the mitochondria independently of Bax activation.
Figure 2: Oxidized cofilin binds to mitochondria, causing their swelling and cytochrome c release through opening of the PTP.
Figure 3: TnCl-induced apoptosis is dependent on oxidation of Cys residues in cofilin, as determined by site-directed mutagenesis and siRNA silencing.
Figure 4: Oxidation of cofilin is required for its translocation to the mitochondria.
Figure 5: Cofilin must be dephosphorylated and oxidized for TnCl to cause apoptosis and loss of cellular mitochondrial membrane potential.

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  • 15 October 2009

    In the version of this letter initially published, the following sentence and reference were ommited: However, a recent study has also reported that cofilin oxidation impairs its cytoskeletal function in T cells28. 28. Klemke, M, Wabnitz, G. H., Funke, F., Funk, B., Kirchgessner, H. & Samstag, Y. Oxidation of cofilin mediates T cell hyporesponsiveness under oxidative stress conditions. Immunity 29, 404-413 (2008). This error has been corrected in the HTML and PDF versions of the letter.

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Acknowledgements

We thank the members of the Laboratory of Biochemistry for critical comments and discussions, and R. Youle and P. Clerc for providing Bax/Bak knockout cells and an anti-Bax N terminus antibody. The research was supported in part by the Brazilian MCT/CNPq Universal funds (479860/2006-8). F.K. and S.Z. were supported by training grants from the Oak Ridge Institute for Science and Education. This project was funded in part by federal funds from the National Cancer Institute, National Institutes of Health, under Contract NO1-CO-12400. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the United States Government. The authors would like to dedicate this manuscript to the memory of Earl R. Stadtman for his mentoring and pioneering work in biochemistry and protein oxidation.

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Author notes

  1. Li-Rong Yu

    Present address: Current address: Center for Proteomics, National Center for Toxicological Research, Food and Drug Administration, Jefferson, AR 72079, USA.,

  2. Fábio Klamt and Stéphanie Zdanov: These authors contributed equally to this work.

Authors and Affiliations

  1. Division of Therapeutic Proteins, Laboratory of Biochemistry, Center for Drug Evaluation and Research, Food and Drug Administration, Bethesda, 20892, MD, USA

    Fábio Klamt, Stéphanie Zdanov, Ashley Pariser, Yaqin Zhang, Baolin Zhang & Emily Shacter

  2. Department of Biochemistry, Center of Oxidative Stress Research, ICBS/Federal University of Rio Grande do Sul (UFRGS), Porto Alegre, 90035-003, RS, Brazil

    Fábio Klamt

  3. Laboratory of Biochemistry, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, 20892, MD, USA

    Rodney L. Levine

  4. Laboratory of Proteomics and Analytical Technologies, Advanced Technologies program, SAIC-Frederick Inc., National Cancer Institute at Frederick, Frederick, 21702–1201, MD, USA

    Li-Rong Yu & Timothy D. Veenstra

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Contributions

F.K., S.Z., R.L.L. and E.S. carried out project planning, experimental design, experimental work and data analysis. F.K. carried out experiments for Figs 1, 2, 4, 5, S1 and S2, and helped write the manuscript. S.Z. carried out experiments for Figs 3, 4, 5 and S3, S4, S5, S6, S7, and helped write the manuscript. R.L.L. carried out experiments for Figs S6 and S7. A.P. carried out experiments and data analysis for Fig. S4. Y.Z. and B.Z. carried out experiments for Fig. 3. L.-R.Y. and T.D.V. carried out experiments and data analysis for Fig. S1. E.S. carried out experiments for Fig. 4, directed the research and wrote the manuscript.

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Correspondence to Emily Shacter.

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Klamt, F., Zdanov, S., Levine, R. et al. Oxidant-induced apoptosis is mediated by oxidation of the actin-regulatory protein cofilin. Nat Cell Biol 11, 1241–1246 (2009). https://doi.org/10.1038/ncb1968

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