Assays for autophagy II: Mitochondrial autophagy.
Autophagy is a bulk degradation process of cytosolic proteins and organelles through the lysosomal/vacuolar machinery. Mitophagy is a type of autophagy that selectively degrades mitochondria. Recent studies have revealed that mitophagy plays an important role in cellular mitochondrial quality control. The budding yeast Saccharomyces cerevisiae is a powerful model that has been applied to study many biological phenomena. This model organism has contributed greatly to our understanding of autophagy, including the identification of more than 30 autophagy-related genes. Similarly, the molecular mechanisms and physiological roles of mitophagy have been gradually elucidated using S. cerevisiae. In this chapter, we describe two commonly used protocols to detect mitophagy in S. cerevisiae: fluorescence microscopy and immunoblotting.
Laboratory of Biosignaling, Graduate School of Medical and Dental Sciences, Niigata University, 1-757 Asahimachi-dori, Chuo-ku, Niigata, 951-8510, Japan, kanki@med.niigata-u.ac.jp.
Tomotake
T
Kanki
Koji
K
Okamoto
D001343:Autophagy; D003600:Cytosol / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000378:metabolism*; D008967:Molecular Biology / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism
D024101:Mitochondrial Proteins