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Ceramide targets autophagosomes to mitochondria and induces lethal mitophagy

Nature Chemical Biology volume 8, pages 831–838 (2012)Cite this article

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An Erratum to this article was published on 26 November 2012

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Abstract

Mechanisms by which autophagy promotes cell survival or death are unclear. We provide evidence that C18-pyridinium ceramide treatment or endogenous C18-ceramide generation by ceramide synthase 1 (CerS1) expression mediates autophagic cell death, independent of apoptosis in human cancer cells. C18-ceramide–induced lethal autophagy was regulated via microtubule-associated protein 1 light chain 3 β-lipidation, forming LC3B-II, and selective targeting of mitochondria by LC3B-II–containing autophagolysosomes (mitophagy) through direct interaction between ceramide and LC3B-II upon Drp1-dependent mitochondrial fission, leading to inhibition of mitochondrial function and oxygen consumption. Accordingly, expression of mutant LC3B with impaired ceramide binding, as predicted by molecular modeling, prevented CerS1-mediated mitochondrial targeting, recovering oxygen consumption. Moreover, knockdown of CerS1 abrogated sodium selenite–induced mitophagy, and stable LC3B knockdown protected against CerS1- and C18-ceramide–dependent mitophagy and blocked tumor suppression in vivo. Thus, these data suggest a new receptor function of ceramide for anchoring LC3B-II autophagolysosomes to mitochondrial membranes, defining a key mechanism for the induction of lethal mitophagy.

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Figure 1: Exogenous C18-Pyr-Cer induces autophagic cell death.
Figure 2: C18-Pyr-Cer mediates targeting of autophagolysosomes to mitochondria and the inhibition of mitochondrial function.
Figure 3: Induction of endogenous C18-ceramide by CerS1 expression mediates lethal mitophagy.
Figure 4: Mitochondrial localization of CerS1-generated C18-ceramide induces targeting of mitochondria by LC3B-II–containing autophagosomes.
Figure 5: C18-ceramide interacts with LC3B via lipid-protein association.
Figure 6: C18-ceramide–LC3B-II interaction on mitochondrial membranes is regulated downstream of Drp1-mediated mitochondrial fission.
Figure 7: The C18-ceramide–LC3B interaction induces lethal mitophagy and subsequent tumor suppression in vivo.

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Change history

  • 11 October 2012

    In the version of this article initially published, the numbering of amino acids Ile35 and Phe52 for LC3B in Figure 5 was incorrect. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We thank J.G. Schnellmann for her editorial review. We also thank D.J. Hazen-Martin and her group for TEM preparations. This work was supported by research grants obtained from the US National Institutes of Health (NIH; CA088932, DE016572 and CA097165 to B.O.). The core facilities used for animal studies, lipidomics and imaging were constructed using support from NIH (C06 RR015455, 5-P30-DK34987, 1-P50-AA11605 and P30 CA138313).

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Author notes

  1. R David Sentelle and Can E Senkal: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, South Carolina, USA

    R David Sentelle, Can E Senkal, Wenhui Jiang, Suriyan Ponnusamy, Salih Gencer, Shanmugam Panneer Selvam, Zdzislaw M Szulc, Jacek Bielawski & Besim Ogretmen

  2. Hollings Cancer Center, Medical University of South Carolina, Charleston, South Carolina, USA

    R David Sentelle, Can E Senkal, Wenhui Jiang, Suriyan Ponnusamy, Salih Gencer, Shanmugam Panneer Selvam, Venkat K Ramshesh, Yuri K Peterson, John J Lemasters, Zdzislaw M Szulc, Jacek Bielawski & Besim Ogretmen

  3. Department of Pharmaceutical Sciences, Medical University of South Carolina, Charleston, South Carolina, USA

    Venkat K Ramshesh, Yuri K Peterson & John J Lemasters

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Contributions

R.D.S. performed mitophagy analyses, cell death and OCR measurements and xenograft studies; C.E.S. performed ceramide–LC3B-II binding assays and confocal microscopy; W.J. performed confocal microscopy and OCR measurements; S.P. measured LC3B lipidation; S.G. measured cellular ATP; S.P.S. measured LC3B lipidation in MEFs; V.K.R. performed immunofluorescence and confocal microscopy; Y.K.P. performed the structural analysis and docking simulations; J.J.L. designed experiments for detection of mitophagy by confocal microscopy and analyzed data; Z.M.S. designed and synthesized ceramide analogues; J.B. measured ceramides using lipidomics; and B.O. conceived and designed experiments, analyzed data and wrote the manuscript.

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Correspondence to Besim Ogretmen.

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Sentelle, R., Senkal, C., Jiang, W. et al. Ceramide targets autophagosomes to mitochondria and induces lethal mitophagy. Nat Chem Biol 8, 831–838 (2012). https://doi.org/10.1038/nchembio.1059

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