TY - JOUR
T1 - Fluorescence microscopy-based assays for monitoring yeast Atg protein trafficking.
AU - Shintani, Takahiro
AU - Reggiori, Fulvio
N1 - Funding Information:
The authors thank Aniek van der Vaart and Muriel Mari for the critical reading of the manuscript. T.S. is supported by Grants‐in‐Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (19580391). F.R. is supported by the Netherlands Organization for Health Research and Development (ZonMW‐VIDI‐917.76.329) and by the Utrecht University (High Potential grant).
PY - 2008
Y1 - 2008
N2 - For several years, the yeast Saccharomyces cerevisiae has been the leading model organism for the study of autophagy. The amenability of this unicellular eukaryote to genetic and biochemical approaches has allowed the isolation and characterization of most of the genes specifically involved in autophagy, which are known as ATG (Reggiori, 2006; Reggiori and Klionsky, 2005). These pioneering studies have been of crucial relevance because most of the yeast ATG genes possess orthologs in all eukaryotic organisms. The experimental advantages, all the available reagents, and the established assays still maintain yeast in a prominent position in the study of autophagy and autophagy-related pathways. In this chapter, we describe fluorescent protein-based methodologies that permit one to readily assay the functionality of the autophagic pathway and to assess the trafficking of one of the key protein of this degradative process, Atg9.
AB - For several years, the yeast Saccharomyces cerevisiae has been the leading model organism for the study of autophagy. The amenability of this unicellular eukaryote to genetic and biochemical approaches has allowed the isolation and characterization of most of the genes specifically involved in autophagy, which are known as ATG (Reggiori, 2006; Reggiori and Klionsky, 2005). These pioneering studies have been of crucial relevance because most of the yeast ATG genes possess orthologs in all eukaryotic organisms. The experimental advantages, all the available reagents, and the established assays still maintain yeast in a prominent position in the study of autophagy and autophagy-related pathways. In this chapter, we describe fluorescent protein-based methodologies that permit one to readily assay the functionality of the autophagic pathway and to assess the trafficking of one of the key protein of this degradative process, Atg9.
UR - http://www.scopus.com/inward/record.url?scp=61949147675&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=61949147675&partnerID=8YFLogxK
M3 - Article
C2 - 19185712
AN - SCOPUS:61949147675
VL - 451
SP - 43
EP - 56
JO - Methods in Enzymology
JF - Methods in Enzymology
SN - 0076-6879
ER -