title: 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.
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: William E
authors.initials: WE
authors.lastname: Dowdle
authors.affiliation: Novartis Institutes for BioMedical Research, Novartis Campus, CH-4056 Basel, Switzerland.
authors.forename: Beat
authors.initials: B
authors.lastname: Nyfeler
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Jane
authors.initials: J
authors.lastname: Nagel
authors.affiliation: Novartis Institutes for BioMedical Research, 4560 Horton Street Emeryville, California 94608, USA.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Elling
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Shanming
authors.initials: S
authors.lastname: Liu
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Ellen
authors.initials: E
authors.lastname: Triantafellow
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Suchithra
authors.initials: S
authors.lastname: Menon
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Zuncai
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Ayako
authors.initials: A
authors.lastname: Honda
authors.affiliation: Novartis Institutes for BioMedical Research, 4560 Horton Street Emeryville, California 94608, USA.
authors.forename: Gwynn
authors.initials: G
authors.lastname: Pardee
authors.affiliation: Novartis Institutes for BioMedical Research, 4560 Horton Street Emeryville, California 94608, USA.
authors.forename: John
authors.initials: J
authors.lastname: Cantwell
authors.affiliation: Novartis Institutes for BioMedical Research, 4560 Horton Street Emeryville, California 94608, USA.
authors.forename: Catherine
authors.initials: C
authors.lastname: Luu
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Ivan
authors.initials: I
authors.lastname: Cornella-Taracido
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Edmund
authors.initials: E
authors.lastname: Harrington
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Fekkes
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Lei
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Qing
authors.initials: Q
authors.lastname: Fang
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Mary Ellen
authors.initials: ME
authors.lastname: Digan
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Debra
authors.initials: D
authors.lastname: Burdick
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Andrew F
authors.initials: AF
authors.lastname: Powers
authors.affiliation: Novartis Institutes for BioMedical Research, Novartis Campus, CH-4056 Basel, Switzerland.
authors.forename: Stephen B
authors.initials: SB
authors.lastname: Helliwell
authors.affiliation: Novartis Institutes for BioMedical Research, Novartis Campus, CH-4056 Basel, Switzerland.
authors.forename: Simon
authors.initials: S
authors.lastname: D'Aquin
authors.affiliation: Novartis Institutes for BioMedical Research, Novartis Campus, CH-4056 Basel, Switzerland.
authors.forename: Julie
authors.initials: J
authors.lastname: Bastien
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Henry
authors.initials: H
authors.lastname: Wang
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Dmitri
authors.initials: D
authors.lastname: Wiederschain
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Jenny
authors.initials: J
authors.lastname: Kuerth
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Philip
authors.initials: P
authors.lastname: Bergman
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: David
authors.initials: D
authors.lastname: Schwalb
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Jason
authors.initials: J
authors.lastname: Thomas
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Savuth
authors.initials: S
authors.lastname: Ugwonali
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Fred
authors.initials: F
authors.lastname: Harbinski
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: John
authors.initials: J
authors.lastname: Tallarico
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Wilson
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Vic E
authors.initials: VE
authors.lastname: Myer
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Jeffery A
authors.initials: JA
authors.lastname: Porter
authors.affiliation: Novartis Institutes for BioMedical Research, 4560 Horton Street Emeryville, California 94608, USA.
authors.forename: Dirksen E
authors.initials: DE
authors.lastname: Bussiere
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Peter M
authors.initials: PM
authors.lastname: Finan
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Labow
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Mao
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Lawrence G
authors.initials: LG
authors.lastname: Hamann
authors.affiliation: Department of Genetics and Complex Diseases, Harvard School of Public Health, 665 Huntington Avenue, Boston, Massachusetts 02115, USA.
authors.forename: Brendan D
authors.initials: BD
authors.lastname: Manning
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Reginald A
authors.initials: RA
authors.lastname: Valdez
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Thomas
authors.initials: T
authors.lastname: Nicholson
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Markus
authors.initials: M
authors.lastname: Schirle
authors.affiliation: Novartis Institutes for BioMedical Research, 4560 Horton Street Emeryville, California 94608, USA.
authors.forename: Mark S
authors.initials: MS
authors.lastname: Knapp
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Erin P
authors.initials: EP
authors.lastname: Keaney
authors.affiliation: Novartis Institutes for BioMedical Research, 250 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
authors.forename: Leon O
authors.initials: LO
authors.lastname: Murphy
mesh_terms: D000818:Animals; D001343:Autophagy / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D058546:Class III Phosphatidylinositol 3-Kinases / Q000037:antagonists & inhibitors*; D005293:Ferritins / Q000378:metabolism*; D006706:Homeostasis / Q000502:physiology*; D006801:Humans; D007501:Iron / Q000378:metabolism*; D008247:Lysosomes / Q000378:metabolism; D051379:Mice; D056918:Nuclear Receptor Coactivators / Q000378:metabolism*; D010588:Phagosomes / Q000378:metabolism; D011485:Protein Binding
chemical_list: C111382:NCOA4 protein, human; C508724:NcoA4 protein, mouse; D056918:Nuclear Receptor Coactivators; D005293:Ferritins; D007501:Iron; D058546:Class III Phosphatidylinositol 3-Kinases