The C-terminal region of ATG101 bridges ULK1 and PtdIns3K complex in autophagy initiation
| Autor: | Juneyoung Jung, Ick Young Kim, Jiyea Kim, Hyun Kyu Song, Seongman Kang, Byeong Won Kim, Jun Hoe Kim, Yunjung Jin, Joungmok Kim, Heesun Cheong |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
Male
Models Molecular 0301 basic medicine Research Paper - Basic Science Protein subunit Vesicular Transport Proteins Autophagy-Related Proteins UVRAG HORMA domain Biology Crystallography X-Ray Vacuolar Sorting Protein VPS15 03 medical and health sciences Sequestosome 1 X-Ray Diffraction Scattering Small Angle Autophagy Tumor Cells Cultured Autophagy-Related Protein-1 Homolog Humans Protein Interaction Domains and Motifs education Molecular Biology Zinc finger education.field_of_study 030102 biochemistry & molecular biology Intracellular Signaling Peptides and Proteins Cell Biology BECN1 Autophagy-related protein 13 Class III Phosphatidylinositol 3-Kinases Cell biology HEK293 Cells 030104 developmental biology Multiprotein Complexes Protein Binding |
| DOI: | 10.6084/m9.figshare.6938576 |
| Popis: | The initiation of macroautophagy/autophagy is tightly regulated by the upstream ULK1 kinase complex, which affects many downstream factors including the PtdIns3K complex. The phosphorylation of the right position at the right time on downstream molecules is governed by proper complex formation. One component of the ULK1 complex, ATG101, known as an accessory protein, is a stabilizer of ATG13 in cells. The WF finger region of ATG101 plays an important role in the recruitment of WIPI1 (WD repeat domain, phosphoinositide interacting protein 1) and ZFYVE1 (zinc finger FYVE-type containing 1). Here, we report that the C-terminal region identified in the structure of the human ATG101-ATG13HORMA complex is responsible for the binding of the PtdIns3K complex. This region adopts a β-strand conformation in free ATG101, but either an α-helix or random coil in our ATG101-ATG13HORMA complex, which protrudes from the core and interacts with other molecules. The C-terminal deletion of ATG101 shows a significant defect in the interaction with PtdIns3K components and subsequently impairs autophagosome formation. This result clearly presents an additional role of ATG101 for bridging the ULK1 and PtdIns3K complexes in the mammalian autophagy process. Abbreviations: ATG: autophagy related; BECN1: beclin 1; GFP: green fluorescent protein; HORMA: Hop1p/Rev7p/MAD2; HsATG13HORMA: HORMA domain of ATG13 from Homo sapiens; KO: knockout; MAD2: mitotic arrest deficient 2 like 1; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; PIK3C3/VPS34: phosphatidylinositol 3-kinase catalytic subunit type 3; PIK3R4/VPS15: phosphoinositide-3-kinase regulatory subunit 4; PtdIns3K: phosphatidylinositol 3-kinase; RB1CC1/FIP200: RB1 inducible coiled-coil 1; SAXS: small-angle X-ray scattering; ScAtg13HORMA: HORMA domain of Atg13 from Sccharomyces cerevisiae; SEC-SAXS: size-exclusion chromatography with small-angle X-ray scattering; SpAtg13HORMA: HORMA domain of Atg13 from Schizosaccharomyces pombe; SQSTM1/p62: sequestosome 1; ULK1: unc51-like autophagy activating kinase 1; UVRAG: UV radiation resistance associated; WIPI1: WD repeat domain: phosphoinositide interacting 1; ZFYVE1/DFCP1: zinc finger FYVE-type containing 1 |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|