title: Autophagy is a major regulator of beta cell insulin homeostasis.
abstract: We studied the role of protein degradation pathways in the regulation of insulin production and secretion and hypothesised that autophagy regulates proinsulin degradation, thereby modulating beta cell function.
  
  
  
  Proinsulin localisation in autophagosomes was demonstrated by confocal and electron microscopy. Autophagy was inhibited by knockdown of autophagy-related (ATG) proteins and using the H(+)-ATPase inhibitor bafilomycin-A1. Proinsulin and insulin content and secretion were assessed in static incubations by ELISA and RIA.
  
  
  
  Confocal and electron microscopy showed proinsulin localised in autophagosomes and lysosomes. Beta-Atg7 (-/-) mice had proinsulin-containing sequestosome 1 (p62 [also known as SQSTM1])(+) aggregates in beta cells, indicating proinsulin is regulated by autophagy in vivo. Short-term bafilomycin-A1 treatment and ATG5/7 knockdown increased steady-state proinsulin and hormone precursor chromogranin A content. ATG5/7 knockdown also increased glucose- and non-fuel-stimulated insulin secretion. Finally, mutated forms of proinsulin that are irreparably misfolded and trapped in the endoplasmic reticulum are more resistant to degradation by autophagy.
  
  
  
  In the beta cell, transport-competent secretory peptide precursors, including proinsulin, are regulated by autophagy, whereas efficient clearance of transport-incompetent mutated forms of proinsulin by alternative degradative pathways may be necessary to avoid beta cell proteotoxicity. Reduction of autophagic degradation of proinsulin increases its residency in the secretory pathway, followed by enhanced secretion in response to stimuli.
authors.affiliation: Endocrinology and Metabolism Service, Department of Medicine, Hadassah-Hebrew University Medical Center, PO Box 12000, Jerusalem, 91120, Israel.
authors.forename: Yael
authors.initials: Y
authors.lastname: Riahi
authors.affiliation: Endocrinology and Metabolism Service, Department of Medicine, Hadassah-Hebrew University Medical Center, PO Box 12000, Jerusalem, 91120, Israel.|Molecular Dermatology Research Group, Unit of Dermatology and Venereology, Department of Medicine, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Jakob D
authors.initials: JD
authors.lastname: Wikstrom
authors.affiliation: Endocrinology and Metabolism Service, Department of Medicine, Hadassah-Hebrew University Medical Center, PO Box 12000, Jerusalem, 91120, Israel.
authors.forename: Etty
authors.initials: E
authors.lastname: Bachar-Wikstrom
authors.affiliation: Endocrinology and Metabolism Service, Department of Medicine, Hadassah-Hebrew University Medical Center, PO Box 12000, Jerusalem, 91120, Israel.
authors.forename: Nava
authors.initials: N
authors.lastname: Polin
authors.affiliation: Endocrinology and Metabolism Service, Department of Medicine, Hadassah-Hebrew University Medical Center, PO Box 12000, Jerusalem, 91120, Israel.
authors.forename: Hava
authors.initials: H
authors.lastname: Zucker
authors.affiliation: Severance Biomedical Research Institute and Department of Internal Medicine, Yonsei University College of Medicine, Seoul, South Korea.
authors.forename: Myung-Shik
authors.initials: MS
authors.lastname: Lee
authors.affiliation: Department of Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, South Korea.
authors.forename: Wenying
authors.initials: W
authors.lastname: Quan
authors.affiliation: Division of Metabolism, Endocrinology and Diabetes, University of Michigan, Ann Arbor, MI, USA.
authors.forename: Leena
authors.initials: L
authors.lastname: Haataja
authors.affiliation: Division of Metabolism, Endocrinology and Diabetes, University of Michigan, Ann Arbor, MI, USA.
authors.forename: Ming
authors.initials: M
authors.lastname: Liu
authors.affiliation: Division of Metabolism, Endocrinology and Diabetes, University of Michigan, Ann Arbor, MI, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Arvan
authors.affiliation: Endocrinology and Metabolism Service, Department of Medicine, Hadassah-Hebrew University Medical Center, PO Box 12000, Jerusalem, 91120, Israel.
authors.forename: Erol
authors.initials: E
authors.lastname: Cerasi
authors.affiliation: Endocrinology and Metabolism Service, Department of Medicine, Hadassah-Hebrew University Medical Center, PO Box 12000, Jerusalem, 91120, Israel. gleib@hadassah.org.il.
authors.forename: Gil
authors.initials: G
authors.lastname: Leibowitz
mesh_terms: D000818:Animals; D001343:Autophagy / Q000235:genetics / Q000502:physiology*; D000071193:Autophagy-Related Protein 7 / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D002460:Cell Line; D006706:Homeostasis / Q000235:genetics / Q000502:physiology; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D050417:Insulin-Secreting Cells / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008854:Microscopy, Electron; D010101:Oxygen Consumption / Q000235:genetics / Q000502:physiology; D034622:RNA Interference / Q000502:physiology
chemical_list: C502661:Atg7 protein, mouse; D007328:Insulin; D000071193:Autophagy-Related Protein 7
keywords: Autophagy; Beta cells; Insulin secretion; Lysosome; Proinsulin; Proteasome; Protein degradation
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