Structural basis for membrane recruitment of ATG16L1 by WIPI2 in Autophagy

Output Details

Preprint May 14, 2021

Autophagy is a cellular process that degrades cytoplasmic cargo by engulfing it in a double membrane vesicle, known as the autophagosome, and delivering it to the lysosome. The ATG12-5-16L1 complex is responsible for conjugating members of the ubiquitin-like ATG8 protein family to phosphatidylethanolamine in the growing autophagosomal membrane, known as the phagophore. ATG12-5-16L1 is recruited to the phagophore by a subset of the phosphatidylinositol 3-phosphate-binding seven bladed beta-propeller WIPI proteins. We determined the crystal structure of WIPI2d in complex with the WIPI2 interacting region (W2IR) of ATG16L1 comprising residues 207-230 at 1.85 Angstrom resolution. The structure shows that the ATG16L1 W2IR adopts an alpha helical conformation and binds in an electropositive and hydrophobic groove between WIPI2 beta-propeller blades 2 and 3. Mutation of residues at the interface reduces or blocks the recruitment of ATG12-5-16L1 and the conjugation of the ATG8 protein LC3B to synthetic membranes. Interface mutants show a decrease in starvation-induced autophagy. Comparisons across the four human WIPIs suggest that WIPI1 and 2 belong to a W2IR-binding subclass responsible for localizing ATG12-5-16L1 and driving ATG8 lipidation, whilst WIPI3 and 4 belong to a second W34IR-binding subclass responsible for localizing ATG2, and so directing lipid supply to the nascent phagophore. The structure provides a framework for understanding the regulatory node connecting two central events in autophagy initiation, the action of the autophagic PI 3-kinase complex on the one hand, and ATG8 lipidation on the other. Article published in eLife on Sept 10, 2021. Initial Preprint doi: 10.1101/2021.05.14.444175 posted on May 14, 2021.
Identifier (DOI)
10.7554/eLife.70372
Tags
  • ATG12-5-16L1
  • Autophagy
  • Crystallization
  • In Vitro
  • Original Research
  • Structural biology
  • WIPI2d

Meet the Authors

  • Lisa Strong, BSc

    Key Personnel: Team Hurley

    University of California, Berkeley

  • User avatar fallback logo

    Chunmei Chang

  • User avatar fallback logo

    Julia Riley, MSc

    Key Personnel: Team Hurley

    University of Pennsylvania

  • User avatar fallback logo

    C Alexander Boecker

  • User avatar fallback logo

    Thomas G. Flower

  • User avatar fallback logo

    Cosmo Z. Buffalo

  • User avatar fallback logo

    Xuefeng Ren

  • Erika Holzbaur, PhD

    Co-PI (Core Leadership): Team Hurley

    University of Pennsylvania

  • James Hurley

    Lead PI (Core Leadership): Team Hurley

    University of California, Berkeley