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Related Experiment Video

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ESCRT-mediated phagophore sealing during mitophagy.

Yan Zhen1,2, Hélène Spangenberg1,2, Michael J Munson1,3

  • 1Centre for Cancer Cell Reprogramming, Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, Montebello, Norway.

Autophagy
|August 2, 2019
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Summary
This summary is machine-generated.

The ESCRT machinery, specifically CHMP4B, is recruited to autophagosomes during starvation-induced autophagy. This machinery is crucial for the final closure of phagophores, a key step in mitophagy.

Keywords:
ESCRTmacroautophagymembrane sealingmitophagyphagophore

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • The endosomal sorting complex required for transport (ESCRT) machinery's role in macroautophagy/autophagy is not fully understood.
  • Previous studies suggest ESCRT inactivation leads to autophagic defects.

Purpose of the Study:

  • To elucidate the precise functions of ESCRT proteins in macroautophagy and mitophagy.
  • To investigate the role of ESCRT-III subunit CHMP4B in autophagosome formation and closure.

Main Methods:

  • Live-cell fluorescence microscopy to observe CHMP4B recruitment to nascent autophagosomes.
  • Correlative light and electron microscopy (CLEM) for high-resolution imaging of mitophagosome formation.
  • Optogenetic closure assay to assess phagophore sealing dynamics.
  • Depletion studies of ESCRT-III subunits (e.g., CHMP2A) to evaluate mitophagy flux.

Main Results:

  • CHMP4B transiently associates with nascent autophagosomes and mitophagosomes.
  • Depletion of CHMP2A significantly prolonged CHMP4B dwell time on autophagosomes.
  • CHMP2A depletion impaired phagophore sealing during mitophagy.
  • ESCRT-III subunit depletion inhibited both PRKN/PARKIN-dependent and -independent mitophagy.

Conclusions:

  • The ESCRT machinery, particularly ESCRT-III, is essential for mediating phagophore closure during mitophagy.
  • Phagophore closure mediated by ESCRT is a critical step for efficient mitophagic flux.