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Methods. 2015 Mar;75:3-12. doi: 10.1016/j.ymeth.2014.12.008. Epub 2014 Dec 16.

The yeast Saccharomyces cerevisiae: an overview of methods to study autophagy progression.

Author information

1
Life Sciences Institute, University of Michigan, Ann Arbor, MI 48109, United States.
2
Department of Cell Biology, Center for Molecular Medicine, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands; Department of Cell Biology, University Medical Center Groningen, A. Deusinglaan 1, 9713 AV Groningen, The Netherlands.
3
Life Sciences Institute, University of Michigan, Ann Arbor, MI 48109, United States. Electronic address: klionsky@umich.edu.

Abstract

Macroautophagy (hereafter autophagy) is a highly evolutionarily conserved process essential for sustaining cellular integrity, homeostasis, and survival. Most eukaryotic cells constitutively undergo autophagy at a low basal level. However, various stimuli, including starvation, organelle deterioration, stress, and pathogen infection, potently upregulate autophagy. The hallmark morphological feature of autophagy is the formation of the double-membrane vesicle known as the autophagosome. In yeast, flux through the pathway culminates in autophagosome-vacuole fusion, and the subsequent degradation of the resulting autophagic bodies and cargo by vacuolar hydrolases, followed by efflux of the breakdown products. Importantly, aberrant autophagy is associated with diverse human pathologies. Thus, there is a need for ongoing work in this area to further understand the cellular factors regulating this process. The field of autophagy research has grown exponentially in recent years, and although numerous model organisms are being used to investigate autophagy, the baker's yeast Saccharomyces cerevisiae remains highly relevant, as there are significant and unique benefits to working with this organism. In this review, we will focus on the current methods available to evaluate and monitor autophagy in S. cerevisiae, which in several cases have also been subsequently exploited in higher eukaryotes.

KEYWORDS:

Atg8; Autophagosome; Mitophagy; PAS; Phagophore; Vacuole

PMID:
25526918
PMCID:
PMC4355233
DOI:
10.1016/j.ymeth.2014.12.008
[Indexed for MEDLINE]
Free PMC Article

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