In situ structural analysis reveals membrane shape transitions during autophagosome formation

Output Details

Preprint May 2, 2022

Published September 19, 2022

Autophagosomes are unique organelles which form de novo as double-membrane vesicles engulfing cytosolic material for destruction. Their biogenesis involves a series of membrane transformations with distinctly shaped intermediates whose ultrastructure is poorly understood. Here, we combine cell biology, correlative cryo-electron tomography (ET) and novel data analysis to reveal the step-by-step structural progression of autophagosome biogenesis at high resolution directly within yeast cells. By mapping individual structures onto a timeline based on geometric features, we uncover dynamic changes in membrane shape and curvature. Moreover, we reveal the organelle interactome of growing autophagosomes, highlighting a polar organization of contact sites between the phagophore and organelles such as the vacuole and the ER. Collectively, these findings have important implications for the contribution of different membrane sources during autophagy and for the forces shaping and driving phagophores towards closure without a templating cargo.
Identifier (DOI)
10.1073/pnas.2209823119
Tags
  • Autophagy
  • Cryo-ET
  • In Situ
  • Membrane
  • Original Research
  • Structural biology

Meet the Authors

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    Anna Bieber, MSc

    Key Personnel: Team Harper

    Max Planck Institute of Biochemistry

  • Cristina Capitanio, MSc

    Key Personnel: Team Harper

    Max Planck Institute of Biochemistry

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    Philipp S. Erdmann

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    Fabian Fiedler

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    Florian Beck

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    Chia-Wei Lee

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    Delong Li

  • Gerhard Hummer, PhD

    Collaborating PI: Team Hurley

    Max Planck Institute of Biophysics

  • Brenda Schulman, PhD

    Co-PI (Core Leadership): Team Harper

    Max Planck Institute of Biochemistry

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    Wolfgang Baumeister

  • Florian Wilfling, PhD

    Collaborating PI: Team Harper Team Hurley

    Max Planck Institute of Biophysics