[en] The cyclin-dependent kinase inhibitor p27Kip1 (p27) has been involved in promoting autophagy and survival in conditions of metabolic stress. While the signaling cascade upstream of p27 leading to its cytoplasmic localization and autophagy induction has been extensively studied, how p27 stimulates the autophagic process remains unclear. Here, we investigated the mechanism by which p27 promotes autophagy upon glucose deprivation. Mouse embryo fibroblasts (MEFs) lacking p27 exhibit a decreased autophagy flux compared to wild-type cells and this is correlated with an abnormal distribution of autophagosomes. Indeed, while autophagosomes are mainly located in the perinuclear area in wild-type cells, they are distributed throughout the cytoplasm in p27-null MEFs. Autophagosome trafficking towards the perinuclear area, where most lysosomes reside, is critical for autophagosome-lysosome fusion and cargo degradation. Vesicle trafficking is mediated by motor proteins, themselves recruited preferentially to acetylated microtubules, and autophagy flux is directly correlated to microtubule acetylation levels. p27-/- MEFs exhibit a marked reduction in microtubule acetylation levels and restoring microtubule acetylation in these cells, either by re-expressing p27 or with deacetylase inhibitors, restores perinuclear positioning of autophagosomes and autophagy flux. Finally, we find that p27 promotes microtubule acetylation by binding to and stabilizing α-tubulin acetyltransferase (ATAT1) in glucose-deprived cells. ATAT1 knockdown results in random distribution of autophagosomes in p27+/+ MEFs and impaired autophagy flux, similar to that observed in p27-/- cells. Overall, in response to glucose starvation, p27 promotes autophagy by facilitating autophagosome trafficking along microtubule tracks by maintaining elevated microtubule acetylation via an ATAT1-dependent mechanism.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Nowosad, Ada ; MCD, Centre de Biologie Intégrative, Université de Toulouse, CNRS, UPS, 31062, Toulouse, Cedex, France
Creff, Justine; MCD, Centre de Biologie Intégrative, Université de Toulouse, CNRS, UPS, 31062, Toulouse, Cedex, France
Jeannot, Pauline; MCD, Centre de Biologie Intégrative, Université de Toulouse, CNRS, UPS, 31062, Toulouse, Cedex, France
Culerrier, Raphael; MCD, Centre de Biologie Intégrative, Université de Toulouse, CNRS, UPS, 31062, Toulouse, Cedex, France
Codogno, Patrice ; Institut Necker-Enfants Malades (INEM), INSERM U1151-CNRS UMR 8253, F-75015, Paris, France ; Université Paris, F-75005, Paris, France
Manenti, Stephane; Cancer Research Center of Toulouse (CRCT), INSERM U1037, CNRS ERL5294, University of Toulouse, Toulouse, France
The authors thank Raphael Culerrier for technical assistance with cloning. We are grateful to Maxence Nachury (Stanford University) and Jayanta Debnath (University of California San Francisco) for providing reagents. A.N. was supported by a studentship from the Fondation ARC pour la Recherche sur le Cancer. S.M. is supported by a grant from the Ligue Nationale Contre le Cancer. This project was supported by funds from the Ligue Nationale Contre le Cancer (# 2FI13911UPQV) and Fondation ARC pour la Recherche sur le Cancer (PJA 20171206148) to A.B. A.B. is supported by funds from the Fondation pour la Recherche Médicale (FRM Equipe EQU202103012639).A.N. was supported by a studentship from the Fondation ARC pour la Recherche sur le Cancer. S.M. is supported by a grant from the Ligue Nationale Contre le Cancer. This project was supported by funds from the Ligue Nationale Contre le Cancer (# 2FI13911UPQV) and Fondation ARC pour la Recherche sur le Cancer (PJA 20171206148) to A.B. A.B. is supported by funds from the Fondation pour la Recherche Médicale (FRM Equipe EQU202103012639).
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