title: Monitoring the Formation of Autophagosomal Precursor Structures in Yeast Saccharomyces cerevisiae.
abstract: The budding yeast Saccharomyces cerevisiae is a powerful and versatile model organism for studying multiple aspects of the biology of eukaryotic cells, including the molecular principles underlying autophagy. One of the unique advantages of this unicellular system is its amenability to genetic and biochemical approaches, which had a pivotal role in the discovery and characterization of most of the autophagy-related (Atg) proteins, the central players of autophagy. The relevance of investigating autophagy in this cell model lies in the high conservation of this pathway among eukaryotes, i.e., most of the yeast Atg proteins possess one or more mammalian orthologs. In addition to the experimental advantages, a very large collection of reagents keeps S. cerevisiae in a leading position for the study of the molecular mechanism and regulation of autophagy. In this chapter, we describe fluorescence microscopy and biochemical methods that allow to monitor in vivo the assembly the of Atg machinery, a key step of autophagy. These approaches can be very useful to those researchers that would like to assess the progression of the autophagosomal precursor structure formation under various conditions, in the presence of specific Atg protein mutants or in the absence of other factors.
authors.affiliation: University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: R
authors.initials: R
authors.lastname: Gómez-Sánchez
authors.affiliation: University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: J
authors.initials: J
authors.lastname: Sánchez-Wandelmer
authors.affiliation: University of Groningen, University Medical Center Groningen, Groningen, The Netherlands. Electronic address: f.m.reggiori@umcg.nl.
authors.forename: F
authors.initials: F
authors.lastname: Reggiori
mesh_terms: D000071182:Autophagosomes / Q000378:metabolism / Q000648:ultrastructure; D001343:Autophagy; D000071183:Autophagy-Related Proteins / Q000032:analysis / Q000378:metabolism; D008856:Microscopy, Fluorescence; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000378:metabolism* / Q000648:ultrastructure; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis / Q000378:metabolism
chemical_list: D000071183:Autophagy-Related Proteins; D029701:Saccharomyces cerevisiae Proteins
keywords: Autophagosome; Autophagy; PAS; Phagophore; Phagophore assembly site; Preautophagosomal structure; Vacuole; Yeast