Characterization of autophagosome formation site by a hierarchical analysis of mammalian Atg proteins
Open accessAbstract
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.
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Two ubiquitin-like conjugation systems contribute to autophagosome formation.
“Two ubiquitin-like conjugation systems also contribute to autophagosome formation [23,24,25].”
From LC3-Associated Phagocytosis (LAP): Connections with Host Autophagy (2012) · cites this paper as [24]
Human WIPI1 and WIPI2 function downstream of the phosphatidylinositol 3-kinase class III (PtdIns3KC3) complex I and upstream of both the ATG12 and LC3 ubiquitin-like conjugation systems.
“Human WIPI1 and WIPI2 function downstream of the phosphatidylinositol 3-kinase class III (PtdIns3KC3) complex I [PIK3C3, BECN1, PIK3R4/ VPS15, ATG14] and upstream of both the ATG12 and LC3 ubiquitin-like conjugation systems.418,420,421”
From Guidelines for the use and interpretation of assays for monitoring autophagy (2012) · cites this paper as [420]
VMP1 is an essential autophagy gene that is conserved from Dictyostelium to mammals, and the VMP1 protein regulates early steps of the autophagic pathway.
“VMP1 is an essential autophagy gene that is conserved from Dictyostelium to mammals, 232,462 and the VMP1 protein regulates early steps of the autophagic pathway.420”
From Guidelines for the use and interpretation of assays for monitoring autophagy (2012) · cites this paper as [420]
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- Authors
- Itakura, E., & Mizushima, N.
- Affiliations
- Tokyo Medical and Dental University (JP)
- Venue
- Autophagy
- Publisher
- Informa UK Limited
- Published
- 2010-08-16
- Volume
- 6
- Issue
- 6
- Pages
- 764-776
- ISSN
- 1554-8627 (ISSN-L)
- 1554-8635
- PMC
- PMC3321844
- OpenAlex
- W2086096948
- Cited by
- 843
- References
- 6
- Citations by year
- 2026: 28 · 2025: 31 · 2024: 42 · 2023: 49 · 2022: 54 · 2021: 66 · 2020: 61 · 2019: 74 · 2018: 55 · 2017: 53 · 2016: 56 · 2015: 57 · 2014: 64 · 2013: 55 · 2012: 61
- FWCI
- 35.61
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- cc-by-nc
- Locations
- Autophagy (publishedVersion, cc-by-nc)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- University of Michigan
- Takeda Science Foundation
- University of California, San Diego
- Japan Society for the Promotion of Science
- Topics
- Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Keywords
- autophagosome formation, autophagy-related proteins, ULK1, ATG14, FIP200, LC3, PI3-kinase, VMP1, endoplasmic reticulum, starvation, wortmannin, autophagy initiation
- MeSH terms
- Wortmannin, Androstadienes, Animals, Autophagy, Cell Compartmentation, Cell Line, Endoplasmic Reticulum, Humans, Intracellular Membranes, Mammals, Membrane Proteins, Microtubule-Associated Proteins, Models, Biological, Phagosomes, Recombinant Fusion Proteins, Mice, Knockout, Phosphatidylinositol 3-Kinases, Protein Transport, Green Fluorescent Proteins, Mice
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