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Clathrin-coated vesicles use endocytosis to transport receptors and lysosomal hydrolases from the Golgi to the lysosome in the late secretory pathway. Clathrin-mediated endocytosis was the first described endocytic process, and Clathrin-coated vesicles remain one of the most well-studied transport vesicles. The molecular machinery that generates clathrin-coated vesicles comprises over 50 proteins that precisely coordinate vesicle formation. Cell surface receptors concentrated in indented sites...
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Evidence for ESCRT- and clathrin-dependent microautophagy.

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This study reveals the molecular mechanisms of microautophagy in yeast, showing the endosomal sorting complex required for transport (ESCRT) machinery is crucial. It highlights Vps27

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Autophagy Research

Background:

  • Microautophagy is a cellular process where the lysosomal/vacuolar membrane directly engulfs cellular components.
  • The precise molecular machinery governing microautophagy membrane dynamics remains largely unknown.
  • Autophagy, including microautophagy, plays critical roles in cellular homeostasis and stress response.

Purpose of the Study:

  • To elucidate the molecular machinery driving microautophagy in Saccharomyces cerevisiae.
  • To investigate the role of the endosomal sorting complex required for transport (ESCRT) machinery in microautophagy.
  • To understand the mechanism of cargo recognition and engulfment during microautophagy.

Main Methods:

  • Utilized immunochemical monitoring of fluorescently tagged yeast vacuolar transmembrane proteins (Vph1 and Pho8).
  • Observed changes in protein localization following a diauxic shift.
  • Investigated the requirement of ESCRT components, specifically Vps27 and its interaction with clathrin.

Main Results:

  • Demonstrated induction of microautophagy in yeast after a diauxic shift.
  • Identified the ESCRT machinery, including Vps27, as essential for this microautophagy process.
  • Showed Vps27 relocalizes to the vacuolar membrane and its interaction with clathrin is functionally important for lipid droplet uptake.

Conclusions:

  • The ESCRT machinery, particularly Vps27, plays a critical role in mediating microautophagy.
  • This pathway of microautophagy does not depend on the core Atg proteins typically associated with autophagy.
  • The findings provide new insights into the non-canonical mechanisms of autophagy and cellular cargo trafficking.