Selective VPS34 inhibitor blocks autophagy and uncovers a role for NCOA4 in ferritin degradation and iron homeostasis in vivo
Abstract
Cells rely on autophagy to clear misfolded proteins and damaged organelles to maintain cellular homeostasis. In this study we use the new autophagy inhibitor PIK-III to screen for autophagy substrates. PIK-III is a selective inhibitor of VPS34 that binds a unique hydrophobic pocket not present in related kinases such as PI(3)Kα. PIK-III acutely inhibits autophagy and de novo lipidation of LC3, and leads to the stabilization of autophagy substrates. By performing ubiquitin-affinity proteomics on PIK-III-treated cells we identified substrates including NCOA4, which accumulates in ATG7-deficient cells and co-localizes with autolysosomes. NCOA4 directly binds ferritin heavy chain-1 (FTH1) to target the iron-binding ferritin complex with a relative molecular mass of 450,000 to autolysosomes following starvation or iron depletion. Interestingly, Ncoa4(-/-) mice exhibit a profound accumulation of iron in splenic macrophages, which are critical for the reutilization of iron from engulfed red blood cells. Taken together, the results of this study provide a new mechanism for selective autophagy of ferritin and reveal a previously unappreciated role for autophagy and NCOA4 in the control of iron homeostasis in vivo.
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- Authors
- Dowdle, W. E., Nyfeler, B., Nagel, J., Elling, R. A., Liu, S., Triantafellow, E., Menon, S., Wang, Z., Honda, A., Pardee, G., Cantwell, J., Luu, C., Cornella-Taracido, I., Harrington, E., Fekkes, P., Lei, H., Fang, Q., Digan, M. E., Burdick, D., Powers, A. F., Helliwell, S. B., D’Aquin, S., Bastien, J., Wang, H., Wiederschain, D., Kuerth, J., Bergman, P., Schwalb, D., Thomas, J., Ugwonali, S., Harbinski, F., Tallarico, J., Wilson, C. J., Myer, V. E., Porter, J. A., Bussiere, D. E., Finan, P. M., Labow, M. A., Mao, X., Hamann, L. G., Manning, B. D., Valdez, R. A., Nicholson, T., Schirle, M., Knapp, M. S., Keaney, E. P., & Murphy, L. O.
- Affiliations
- Novartis (Switzerland) (CH)
- Harvard University (US)
- Corresponding author
- Murphy, Leon O.
- ORCID
- Dowdle, William E.
- Nyfeler, Beat
- Menon, Suchithra
- Honda, Ayako
- Luu, Catherine
- Harrington, Edmund
- Helliwell, Stephen B.
- Wiederschain, Dmitri
- Thomas, Jason
- Tallarico, John
- Wilson, Christopher J.
- Myer, Vic E.
- Bussiere, Dirksen E.
- Hamann, Lawrence G.
- Manning, Brendan D.
- Valdez, Reginald A.
- Nicholson, Thomas
- Schirle, Markus
- Knapp, Mark S.
- Venue
- Nature Cell Biology
- Published
- 2014-10-19
- Volume
- 16
- Issue
- 11
- Pages
- 1069-1079
- ISSN
- 1465-7392 (ISSN-L)
- 1476-4679
- OpenAlex
- W2013544053
- Cited by
- 759
- References
- 12
- Citations by year
- 2026: 61 · 2025: 99 · 2024: 78 · 2023: 78 · 2022: 81 · 2021: 68 · 2020: 63 · 2019: 51 · 2018: 47 · 2017: 47 · 2016: 49 · 2015: 33 · 2014: 3
- FWCI
- 33.59
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Cell Biology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
- Health Sciences › Medicine › Hematology › Iron Metabolism and Disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Keywords
- Vps34, autophagy inhibition, NCOA4, ferritin degradation, iron homeostasis, FTH1, autolysosomes, ATG7, LC3 lipidation, splenic macrophages
- MeSH terms
- Animals, Autophagy, Cells, Cultured, Ferritins, Homeostasis, Humans, Iron, Lysosomes, Phagosomes, Protein Binding, Mice, Nuclear Receptor Coactivators, Class III Phosphatidylinositol 3-Kinases
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