Assays for autophagy I: The cvt pathway and nonselective autophagy

Abstract

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.