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October 25, 1993FEBS Letters1,750 citationsOpen Access

Isolation and characterization of autophagy‐defective mutants of Saccharomyces cerevisiae

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MTMiki TsukadaYOYoshinori Ohsumi

Key Points

  • The aim is to isolate and characterize autophagy-defective mutants in Saccharomyces cerevisiae to understand their role in protein degradation.
  • Isolation of apg1 mutant via light microscopy from a mutagenized proteinase-deficient strain.
  • Screening and selection of 75 additional apg mutants based on loss of viability during nitrogen starvation.
  • Genetic analysis revealed single recessive chromosomal mutations in the apg mutants.
  • The apg1 mutant showed faster loss of viability compared to wild-type during nitrogen starvation.
  • Fifteen complementation groups were identified among the apg mutants, indicating diverse genetic involvement.
  • All apg mutants were defective in protein degradation induced by nitrogen starvation, demonstrating autophagy's necessity.

Abstract

Autophagy in the yeast is similar to that in mammalian cells. A mutant designated as apg1 (autophagy) defective in accumulation of autophagic bodies in the vacuoles was isolated by selection using a light microscope from a mutagenized proteinase-deficient strain. In the apg1 strain, which has normal vacuolar proteinases, nitrogen starvation did not induce protein degradation. The apg1 mutant lost its viability faster than wild-type cells during nitrogen starvation. By using the loss of viability as a first screening test, 75 other apg mutants were selected. These apg mutants including apg1 fell into 15 complementation groups. Genetic analyses of representative apg mutants revealed that they all had single recessive chromosomal mutations. Strains with each apg mutation were defective in protein degradation in the vacuoles induced by nitrogen starvation and homozygous diploids for each apg mutation did not sporulate. These results on the apg mutants suggest that autophagy via autophagic bodies is indispensable for protein degradation in the vacuoles under starvation conditions, and that at least 15 APG genes are involved in autophagy in yeast.

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Cite This Study

Tsukada et al. (1993) studied this question.

synapsesocial.com/papers/69dd3291a6ead7ee0940c602https://doi.org/10.1016/0014-5793(93)80398-e
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