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Ceramide targets autophagosomes to mitochondria and induces lethal mitophagy

An Erratum to this article was published on 26 November 2012

This article has been updated

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.

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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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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