Assays for autophagy I: the Cvt pathway and nonselective autophagy.
Autophagy is a major intracellular degradation pathway. It is responsible for the bulk removal of obsolete or damaged cytoplasmic components, including both soluble proteins and membrane-bound organelles. As a fundamental function of eukaryotic cells, autophagy plays a key role in protecting the cells from stressful conditions. Budding yeast Saccharomyces cerevisiae has been the pioneering model system in autophagy-related research. In this chapter, we describe three basic assays of autophagy in S. cerevisiae: the Ape1 maturation assay, the GFP-Atg8 processing assay, and the Pho8Δ60 assay. These assays cover the selective cytoplasm to vacuole targeting (Cvt) pathway and starvation-induced nonselective autophagy.
Department of Life Science, College of Life Science, National Taiwan University, Taipei, Republic of China.
Wei-Pang
WP
Huang
Takahiro
T
Shintani
Zhiping
Z
Xie
D001343:Autophagy / Q000235:genetics*; D003593:Cytoplasm / Q000378:metabolism*; D008967:Molecular Biology / Q000379:methods*; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics*; D014617:Vacuoles / Q000378:metabolism