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PMID: 20639694 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Characterization of autophagosome formation site by a hierarchical analysis of mammalian Atg proteins.

Autophagy ·Vol. 6 ·No. 6 ·2010-08-00 ·Pages 764-76

Itakura E, Mizushima N

Abstract

Autophagy is an intracellular degradation process, through which cytosolic materials are delivered to the lysosome.Despite recent identification of many autophagy-related genes, how autophagosomes are generated remains unclear.Here, we examined the hierarchical relationships among mammalian Atg proteins. Under starvation conditions, ULK1,Atg14, WIPI-1, LC3 and Atg16L1 target to the same compartment, whereas DFCP1 localizes adjacently to these Atgproteins. In terms of puncta formation, the protein complex including ULK1 and FIP200 is the most upstream unit and is required for puncta formation of the Atg14-containing PI3-kinase complex. Puncta formation of both DFCP1 and WIPI-1 requires FIP200 and Atg14. The Atg12-Atg5-Atg16L1 complex and LC3 are downstream units among these factors. The punctate structures containing upstream Atg proteins such as ULK1 and Atg14 tightly associate with the ER, where the ER protein vacuole membrane protein 1 (VMP1) also transiently localizes. These structures are formed even when cells are treated with wortmannin to suppress autophagosome formation. These hierarchical analyses suggest that ULK1, Atg14 and VMP1 localize to the ER-associated autophagosome formation sites in a PI3-kinase activity-independent manner.

MeSH Terms
Androstadienes/pharmacology Animals Autophagy/drug effects Cell Compartmentation/drug effects Cell Line Endoplasmic Reticulum/drug effects,metabolism Green Fluorescent Proteins/metabolism Humans Intracellular Membranes/drug effects,metabolism Mammals/metabolism Membrane Proteins/metabolism Mice Mice, Knockout Microtubule-Associated Proteins/metabolism Models, Biological Phagosomes/drug effects,enzymology,metabolism Phosphatidylinositol 3-Kinases/metabolism Protein Transport/drug effects Recombinant Fusion Proteins/metabolism Wortmannin
Chemicals
Androstadienes Map1lc3b protein, mouse Membrane Proteins Microtubule-Associated Proteins Recombinant Fusion Proteins Green Fluorescent Proteins Wortmannin
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Itakura Eisuke
Department of Physiology and Cell Biology, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo, Japan.
Mizushima Noboru
References (58)
58 references, click to expand
  1. Characterization of the isolation membranes and the limiting membranes of autophagosomes in rat hepatocytes by lectin cytochemistry.
    J Histochem Cytochem. 1990 Apr;38(4):573-80 PMID: 2319125
  2. Functional and physical interaction between Bcl-X(L) and a BH3-like domain in Beclin-1.
    EMBO J. 2007 May 16;26(10):2527-39 PMID: 17446862
  3. WIPI-1alpha (WIPI49), a member of the novel 7-bladed WIPI protein family, is aberrantly expressed in human cancer and is linked to starvation-induced autophagy.
    Oncogene. 2004 Dec 16;23(58):9314-25 PMID: 15602573
  4. Autophagosome requires specific early Sec proteins for its formation and NSF/SNARE for vacuolar fusion.
    Mol Biol Cell. 2001 Nov;12(11):3690-702 PMID: 11694599
  5. Regulation of autophagy by the inositol trisphosphate receptor.
    Cell Death Differ. 2007 May;14(5):1029-39 PMID: 17256008
  6. Autophagosome formation: core machinery and adaptations.
    Nat Cell Biol. 2007 Oct;9(10):1102-9 PMID: 17909521
  7. ULK-Atg13-FIP200 complexes mediate mTOR signaling to the autophagy machinery.
    Mol Biol Cell. 2009 Apr;20(7):1992-2003 PMID: 19225151
  8. The role of autophagy in mammalian development: cell makeover rather than cell death.
    Dev Cell. 2008 Sep;15(3):344-357 PMID: 18804433
  9. Identification of Barkor as a mammalian autophagy-specific factor for Beclin 1 and class III phosphatidylinositol 3-kinase.
    Proc Natl Acad Sci U S A. 2008 Dec 9;105(49):19211-6 PMID: 19050071
  10. Role of the Apg12 conjugation system in mammalian autophagy.
    Int J Biochem Cell Biol. 2003 May;35(5):553-61 PMID: 12672448
  11. Characterization of rat TOM70 as a receptor of the preprotein translocase of the mitochondrial outer membrane.
    J Cell Sci. 2002 May 1;115(Pt 9):1895-905 PMID: 11956321
  12. Distribution of protein disulfide isomerase in rat hepatocytes.
    J Histochem Cytochem. 1988 Dec;36(12):1533-42 PMID: 3192937
  13. Convergence of multiple autophagy and cytoplasm to vacuole targeting components to a perivacuolar membrane compartment prior to de novo vesicle formation.
    J Biol Chem. 2002 Jan 4;277(1):763-73 PMID: 11675395
  14. Autophagy in the epithelial cells of murine seminal vesicle in vitro. Formation of large sheets of nascent isolation membranes, sequestration of the nucleus and inhibition by wortmannin and 3-ethyladenine.
    Cell Tissue Res. 2000 Nov;302(2):253-61 PMID: 11131136
  15. JNK1-mediated phosphorylation of Bcl-2 regulates starvation-induced autophagy.
    Mol Cell. 2008 Jun 20;30(6):678-88 PMID: 18570871
  16. ULK1.ATG13.FIP200 complex mediates mTOR signaling and is essential for autophagy.
    J Biol Chem. 2009 May 1;284(18):12297-305 PMID: 19258318
  17. Two lysosomal membrane proteins, LGP85 and LGP107, are delivered to late endosomes/lysosomes through different intracellular routes after exiting from the trans-Golgi network.
    Biochem Biophys Res Commun. 2003 Feb 21;301(4):833-40 PMID: 12589788
  18. A subdomain of the endoplasmic reticulum forms a cradle for autophagosome formation.
    Nat Cell Biol. 2009 Dec;11(12):1433-7 PMID: 19898463
  19. The role of autophagy during the early neonatal starvation period.
    Nature. 2004 Dec 23;432(7020):1032-6 PMID: 15525940
  20. Vesicular trafficking and autophagosome formation.
    Cell Death Differ. 2009 Jul;16(7):956-65 PMID: 19373247
  21. The pancreatitis-induced vacuole membrane protein 1 triggers autophagy in mammalian cells.
    J Biol Chem. 2007 Dec 21;282(51):37124-33 PMID: 17940279
  22. 3D tomography reveals connections between the phagophore and endoplasmic reticulum.
    Autophagy. 2009 Nov;5(8):1180-5 PMID: 19855179
  23. The roles of intracellular protein-degradation pathways in neurodegeneration.
    Nature. 2006 Oct 19;443(7113):780-6 PMID: 17051204
  24. Autophagosome formation from membrane compartments enriched in phosphatidylinositol 3-phosphate and dynamically connected to the endoplasmic reticulum.
    J Cell Biol. 2008 Aug 25;182(4):685-701 PMID: 18725538
  25. Hierarchy of Atg proteins in pre-autophagosomal structure organization.
    Genes Cells. 2007 Feb;12(2):209-18 PMID: 17295840
  26. Autophagy: process and function.
    Genes Dev. 2007 Nov 15;21(22):2861-73 PMID: 18006683
  27. ATG genes involved in non-selective autophagy are conserved from yeast to man, but the selective Cvt and pexophagy pathways also require organism-specific genes.
    Autophagy. 2007 Mar-Apr;3(2):106-16 PMID: 17204848
  28. Role of FIP200 in cardiac and liver development and its regulation of TNFalpha and TSC-mTOR signaling pathways.
    J Cell Biol. 2006 Oct 9;175(1):121-33 PMID: 17015619
  29. Autophagy fights disease through cellular self-digestion.
    Nature. 2008 Feb 28;451(7182):1069-75 PMID: 18305538
  30. The Atg8 conjugation system is indispensable for proper development of autophagic isolation membranes in mice.
    Mol Biol Cell. 2008 Nov;19(11):4762-75 PMID: 18768753
  31. A novel, human Atg13 binding protein, Atg101, interacts with ULK1 and is essential for macroautophagy.
    Autophagy. 2009 Jul;5(5):649-62 PMID: 19287211
  32. Bcl-2 antiapoptotic proteins inhibit Beclin 1-dependent autophagy.
    Cell. 2005 Sep 23;122(6):927-39 PMID: 16179260
  33. Maturation of autophagic vacuoles in Mammalian cells.
    Autophagy. 2005 Apr;1(1):1-10 PMID: 16874026
  34. Sequestration revisited: integrating traditional electron microscopy, de novo assembly and new results.
    Autophagy. 2007 Nov-Dec;3(6):655-62 PMID: 17603297
  35. p62/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death.
    J Cell Biol. 2005 Nov 21;171(4):603-14 PMID: 16286508
  36. Vacuole membrane protein 1 is an endoplasmic reticulum protein required for organelle biogenesis, protein secretion, and development.
    Mol Biol Cell. 2008 Aug;19(8):3442-53 PMID: 18550798
  37. FIP200, a ULK-interacting protein, is required for autophagosome formation in mammalian cells.
    J Cell Biol. 2008 May 5;181(3):497-510 PMID: 18443221
  38. Molecular machinery of autophagosome formation in yeast, Saccharomyces cerevisiae.
    FEBS Lett. 2007 May 22;581(11):2156-61 PMID: 17382324
  39. Early stages of the secretory pathway, but not endosomes, are required for Cvt vesicle and autophagosome assembly in Saccharomyces cerevisiae.
    Mol Biol Cell. 2004 May;15(5):2189-204 PMID: 15004240
  40. Distinct regulation of autophagic activity by Atg14L and Rubicon associated with Beclin 1-phosphatidylinositol-3-kinase complex.
    Nat Cell Biol. 2009 Apr;11(4):468-76 PMID: 19270693
  41. Multiple regulatory and effector roles of autophagy in immunity.
    Curr Opin Immunol. 2009 Feb;21(1):53-62 PMID: 19269148
  42. Beclin 1 forms two distinct phosphatidylinositol 3-kinase complexes with mammalian Atg14 and UVRAG.
    Mol Biol Cell. 2008 Dec;19(12):5360-72 PMID: 18843052
  43. Kinase-inactivated ULK proteins inhibit autophagy via their conserved C-terminal domains using an Atg13-independent mechanism.
    Mol Cell Biol. 2009 Jan;29(1):157-71 PMID: 18936157
  44. A monomeric red fluorescent protein.
    Proc Natl Acad Sci U S A. 2002 Jun 11;99(12):7877-82 PMID: 12060735
  45. The pre-autophagosomal structure organized by concerted functions of APG genes is essential for autophagosome formation.
    EMBO J. 2001 Nov 1;20(21):5971-81 PMID: 11689437
  46. The early secretory pathway contributes to autophagy in yeast.
    Cell Struct Funct. 2003 Feb;28(1):49-54 PMID: 12655150
  47. Mouse Apg16L, a novel WD-repeat protein, targets to the autophagic isolation membrane with the Apg12-Apg5 conjugate.
    J Cell Sci. 2003 May 1;116(Pt 9):1679-88 PMID: 12665549
  48. Cloning and expression of the rat vacuole membrane protein 1 (VMP1), a new gene activated in pancreas with acute pancreatitis, which promotes vacuole formation.
    Biochem Biophys Res Commun. 2002 Jan 18;290(2):641-9 PMID: 11785947
  49. Evolution of Atg1 function and regulation.
    Autophagy. 2009 Aug;5(6):758-65 PMID: 19411825
  50. Yeast autophagosomes: de novo formation of a membrane structure.
    Trends Cell Biol. 2002 May;12(5):231-5 PMID: 12062171
  51. LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing.
    EMBO J. 2000 Nov 1;19(21):5720-8 PMID: 11060023
  52. Two Beclin 1-binding proteins, Atg14L and Rubicon, reciprocally regulate autophagy at different stages.
    Nat Cell Biol. 2009 Apr;11(4):385-96 PMID: 19270696
  53. Dissection of autophagosome formation using Apg5-deficient mouse embryonic stem cells.
    J Cell Biol. 2001 Feb 19;152(4):657-68 PMID: 11266458
  54. Autophagy dysfunction and ubiquitin-positive protein aggregates in Dictyostelium cells lacking Vmp1.
    Autophagy. 2010 Jan;6(1):100-9 PMID: 20009561
  55. Atg101, a novel mammalian autophagy protein interacting with Atg13.
    Autophagy. 2009 Oct;5(7):973-9 PMID: 19597335
  56. Studies on the mechanisms of autophagy: formation of the autophagic vacuole.
    J Cell Biol. 1990 Jun;110(6):1923-33 PMID: 2351689
  57. Nutrient-dependent mTORC1 association with the ULK1-Atg13-FIP200 complex required for autophagy.
    Mol Biol Cell. 2009 Apr;20(7):1981-91 PMID: 19211835
  58. Autophagy genes in immunity.
    Nat Immunol. 2009 May;10(5):461-70 PMID: 19381141
Article Info
Journal
Autophagy
Abbr.
Autophagy
ISSN
1554-8635
DOI
10.4161/auto.6.6.12709
Published
2010-08-00
Pages
764-76
Language
English
Region
United States
NLM ID
101265188
PMCID
PMC3321844
Subset
IM
External Links
PubMed source DOI: 10.4161/auto.6.6.12709 PMC Full Text View journal details
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