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FIP200, a ULK-interacting protein, is required for autophagosome formation in mammalian cells
Abstract
Autophagy is a membrane-mediated intracellular degradation system. The serine/threonine kinase Atg1 plays an essential role in autophagosome formation. However, the role of the mammalian Atg1 homologues UNC-51–like kinase (ULK) 1 and 2 are not yet well understood. We found that murine ULK1 and 2 localized to autophagic isolation membrane under starvation conditions. Kinase-dead alleles of ULK1 and 2 exerted a dominant-negative effect on autophagosome formation, suggesting that ULK kinase activity is important for autophagy. We next screened for ULK binding proteins and identified the focal adhesion kinase family interacting protein of 200 kD (FIP200), which regulates diverse cellular functions such as cell size, proliferation, and migration. We found that FIP200 was redistributed from the cytoplasm to the isolation membrane under starvation conditions. In FIP200-deficient cells, autophagy induction by various treatments was abolished, and both stability and phosphorylation of ULK1 were impaired. These results suggest that FIP200 is a novel mammalian autophagy factor that functions together with ULKs.
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ULK1 autophosphorylates and transphosphorylates its binding partners ATG13, RB1CC1, and ATG101.
“that ULK1 autophosphorylates and transphosphorylates its binding partners ATG13, RB1CC1, and ATG101 [644, 645, 692-700].”
From Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition) 1 (2021) · cites this paper as [699]
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“…numerous autophagic vacuoles, which occupied 3% of the total cytoplasmic volume ( 2% autophagosomes To monitor the autophagic ability of FIP200 / MEFs more precisely, we determined the autophagy fl ux in these cells by the LC3 turnover assay ( Tanida et al., 2005 ; Mizushima and Yoshimori, 2007 ).”
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Metadata
- Authors
- Hara, T., Takamura, A., Kishi, C., Iemura, S.-I., Natsume, T., Guan, J.-L., & Mizushima, N.
- Affiliations
- 1Department of Physiology and Cell Biology, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo 113-8549, Japan
- 2Biological Information Research Center, National Institutes of Advanced Industrial Science and Technology, Kohtoh-ku, Tokyo 135-0064, Japan
- 3Department of Internal Medicine-MMG, University of Michigan Medical School, Ann Arbor, MI 48109
- Venue
- The Journal of Cell Biology
- Publisher
- Rockefeller University Press
- Published
- 2008-04-28
- Volume
- 181
- Issue
- 3
- Pages
- 497-510
- ISSN
- 1540-8140
- 0021-9525
- PMC
- PMC2364687
- Cited by
- 904
- References
- 10
- Language
- en
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