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Related Experiment Videos

A protein conjugation system essential for autophagy

N Mizushima1, T Noda, T Yoshimori

  • 1Department of Cell Biology, National Institute for Basic Biology, Okazaki, Japan.

Nature
|October 6, 1998
PubMed
Summary

Researchers identified a novel ubiquitination-like conjugation system essential for autophagy in yeast. This system involves Apg12 conjugation to Apg5, mediated by an Apg7 enzyme, and is conserved in mammals.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Autophagy is a critical cellular process for degrading proteins and cytoplasmic components via autophagosomes and lysosomes.
  • This process is vital for cell survival during starvation and for cell differentiation.
  • No specific molecules involved in autophagy had been identified in higher eukaryotes prior to this study.

Purpose of the Study:

  • To investigate the molecular mechanisms underlying autophagy in the yeast Saccharomyces cerevisiae.
  • To identify and characterize molecules essential for the autophagic process.

Main Methods:

  • Isolation and analysis of 14 autophagy-defective (apg) mutants in yeast.
  • Examination of the Apg5/Apg12 protein conjugation system.

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  • Cloning of the APG7 gene and characterization of the Apg7 protein.
  • In vitro reconstitution of the conjugation reaction.

    • Main Results:

    • A unique covalent-modification system, involving the conjugation of Apg12 to Apg5, is essential for autophagy.
    • Mutants apg7 and apg10 lack the Apg5/Apg12 conjugate, indicating their role in this pathway.
    • Apg7 functions as a ubiquitin-E1-like enzyme, facilitating the conjugation.
    • The conjugation reaction is ATP-dependent and can be reconstituted in vitro.
    • Homologues of Apg5 and Apg12 exist in mammalian cells, suggesting conserved mechanisms.

    Conclusions:

    • A novel ubiquitination-like conjugation system, independent of ubiquitin itself, is crucial for autophagy.
    • This modification system, involving Apg7, Apg12, and Apg5, is conserved across species from yeast to mammals.
    • The findings provide the first molecular insight into a key pathway regulating autophagy.