title: Assays for Mitophagy in Yeast.
abstract: Elimination of damaged or surplus mitochondria is crucial to maintain cellular integrity and an energy supply-demand balance. Mitophagy serves to selectively catabolize mitochondria in a manner dependent on autophagy, and contributes to mitochondrial quality and quantity control. This degradation system is highly conserved among eukaryotes including the budding yeast, Saccharomyces cerevisiae. Therefore, analyses of mitophagy using yeast have the potential to provide insights into the common mechanisms of mitophagy. Here, we introduce experimental approaches with fluorescence microscopy and western blotting to validate mitophagy in yeast. The former is useful to visualize transport of mitochondria to the vacuole, a lytic compartment, and formation of mitochondria-containing autophagosomes. The latter allows us to quantify mitochondria degradation.
authors.affiliation: Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, 565-0871, Japan.
authors.forename: Akinori
authors.initials: A
authors.lastname: Eiyama
authors.affiliation: Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, 565-0871, Japan. kokamoto@fbs.osaka-u.ac.jp.
authors.forename: Koji
authors.initials: K
authors.lastname: Okamoto
mesh_terms: D001681:Biological Assay / Q000379:methods*; D015153:Blotting, Western; D017930:Genes, Reporter; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000235:genetics / Q000378:metabolism*; D063306:Mitophagy; D057054:Molecular Imaging / Q000379:methods; D010588:Phagosomes; D014170:Transformation, Genetic; D015003:Yeasts / Q000235:genetics / Q000378:metabolism*
keywords: Mitochondria degradation; Mitophagosome; Mitophagy; Respiration; Yeast