Membrane curvature sensing and stabilization by the autophagic LC3 lipidation machinery

Output Details

Preprint May 4, 2022

Published December 14, 2022

How the highly curved phagophore membrane is stabilized during autophagy initiation is a major open question in autophagosome biogenesis. Here, we use in vitro reconstitution on membrane nanotubes and molecular dynamics simulations to investigate how core autophagy proteins in the LC3 lipidation cascade interact with curved membranes, providing insight into possible roles in regulating membrane shape during autophagosome biogenesis. ATG12–5-16L1 was up to 100-fold enriched on highly curved nanotubes relative to flat membranes. At high surface density, ATG12–5-16L1 binding increased the curvature of the nanotubes. While WIPI2 binding directs membrane recruitment, the amphipathic helix α2 of ATG16L1 is responsible for curvature sensitivity. Molecular dynamics simulations revealed that helix α2 of ATG16L1 inserts shallowly into the membrane, explaining its curvature-sensitive binding to the membrane. These observations show how the binding of the ATG12–5-16L1 complex to the early phagophore rim could stabilize membrane curvature and facilitate autophagosome growth.
Identifier (DOI)
10.1126/sciadv.add1436
Tags
  • ATG12-5-16L1
  • Autophagy
  • In Vitro
  • Membrane curvature
  • Optical Tweezers
  • Original Research
  • WIPI2d

Meet the Authors

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    Liv Jensen

    Key Personnel: Team Hurley

    University of California, Berkeley

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    Shanlin Rao, PhD

    Key Personnel: Team Hurley

    Max Planck Institute of Biophysics

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    Martina Schuschnig

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    A. King Cada

  • Sascha Martens

    Co-PI (Core Leadership): Team Hurley

    Max F. Perutz Laboratories

  • Gerhard Hummer, PhD

    Collaborating PI: Team Hurley

    Max Planck Institute of Biophysics

  • James Hurley

    Lead PI (Core Leadership): Team Hurley

    University of California, Berkeley