Abstract
LC3 is a protein that can associate with autophagosomes, autolysosomes, and phagosomes. Here, we show that LC3 can also redistribute toward the damaged Golgi apparatus where it clusters with SQSTM1/p62 and lysosomes. This organelle-specific relocation, which did not involve the generation of double-membraned autophagosomes, could be observed after Golgi damage was induced by various strategies, namely (i) laser-induced localized cellular damage, (ii) local expression of peroxidase and exposure to peroxide and diaminobenzidine, (iii) treatment with the Golgi-tropic photosensitizer redaporfin and light, (iv) or exposure to the Golgi-tropic anticancer peptidomimetic LTX-401. Mechanistic exploration led to the conclusion that both reactive oxygen species-dependent and -independent Golgi damage induces a similar phenotype that depended on ATG5 yet did not depend on phosphatidylinositol-3-kinase catalytic subunit type 3 and Beclin-1. Interestingly, knockout of ATG5 sensitized cells to Golgi damage-induced cell death, suggesting that the pathway culminating in the relocation of LC3 to the damaged Golgi may have a cytoprotective function.
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Acknowledgements
The plasmid pEGFP-LC3 wt was a gift from Dr Noboru Mizushima (The Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan). The pGalT-EGFP plasmid was a kind gift from J. White, EMBL, Heidelberg, Germany. The pDest-mCherry-p62 plasmid was a kind gift from Terje Johansen. The pExp-Dh1-GFP-LC3-Ef1-Puro for lentiviral production was a kind gift of François-Xavier Gobert and Philippe Benaroch. Luzitin SA provided redaporfin and F2BOH. Sabrina Forveille for her help on assessing colonies numbers. LCGdS is supported by Portuguese Science Foundation (ref. SFRH/BPD/93562/2013). GK is supported by the Ligue contre le Cancer (équipe labelisée); Agence National de la Recherche (ANR)—Projets blancs; ANR under the frame of E-Rare-2, the ERA-Net for Research on Rare Diseases; Association pour la recherche sur le cancer (ARC); Cancéropôle Ile-de-France; Institut National du Cancer (INCa); Institut Universitaire de France; Fondation pour la Recherche Médicale (FDM20140630126 and FDM 40739); the European Commission (ArtForce); the European Research Council (ERC); the LeDucq Foundation; the LabEx Immuno-Oncology; the RHU Torino Lumière, the SIRIC Stratified Oncology Cell DNA Repair and Tumor Immune Elimination (SOCRATE); the SIRIC Cancer Research and Personalized Medicine (CARPEM); and the Paris Alliance of Cancer Research Institutes (PACRI). FP is supported by the Centre National de la Recherche Scientifique (CNRS), the Institut Curie and the LabEx CelTisPhyBio (ANR-10-LBX-0038 part of the IDEX PSL no ANR-10-IDEX-0001–02).
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L.C.G.d.S. performed the experiments with redaporfin-PDT and LTX-401 and A.J.J. performed the experiments of Golgi damage using laser photodamage and HRP-ManII-expressing cells. A.S. developed R scripts used for analysis. S.S., S.D., and M.S. performed electron microscopy, Ø.R. and B.S. provided LTX-401. L.A., O.K., G.K., and F.P. designed the study. G.K. and F.P. wrote the paper.
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BS and ØR are full-time employees of Lytix Biopharma and this study was partially funded by Lytix Biopharma.
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Gomes-da-Silva, L.C., Jimenez, A.J., Sauvat, A. et al. Recruitment of LC3 to damaged Golgi apparatus. Cell Death Differ 26, 1467–1484 (2019). https://doi.org/10.1038/s41418-018-0221-5
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DOI: https://doi.org/10.1038/s41418-018-0221-5
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