title: Identifying a Rab effector on the macroautophagy pathway.
abstract: Rab GTPases are key regulators of membrane traffic. The Rab GTPase Ypt1 is essential for endoplasmic reticulum (ER)-Golgi traffic, intra-Golgi traffic, and the macroautophagy pathway. To identify effectors on the macroautophagy pathway, known autophagy-related genes (Atg genes) required for macroautophagy were tagged with GFP and screened for mislocalization in the ypt1-2 mutant. At the pre-autophagosomal structure (PAS), the localization of the serine/threonine kinase Atg1 was affected in the ypt1-2 mutant. We then used an in vitro binding assay to determine if Atg1 and Ypt1 physically interact with each other and co-immunoprecipitation experiments were performed to address if Atg1 preferentially interacts with the GTP-bound form of Ypt1.
authors: Department of Cellular and Molecular Medicine, Howard Hughes Medical Institute, University of California at San Diego, George Palade Labs, Room 315, La Jolla, CA, 92093-0668, USA.
authors: Juan
authors: J
authors: Wang
authors: Serena
authors: S
authors: Cervantes
authors: Saralin
authors: S
authors: Davis
authors: Susan
authors: S
authors: Ferro-Novick
mesh_terms: D001343:Autophagy; D000071183:Autophagy-Related Proteins; D005978:Glutathione / Q000737:chemistry; D006160:Guanosine Triphosphate / Q000378:metabolism; D047468:Immunoprecipitation; D008856:Microscopy, Fluorescence; D009154:Mutation; D010588:Phagosomes / Q000378:metabolism; D025941:Protein Interaction Mapping / Q000379:methods*; D011494:Protein Kinases / Q000302:isolation & purification / Q000378:metabolism; D021381:Protein Transport; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012685:Sepharose / Q000737:chemistry; D020691:rab GTP-Binding Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D000071183:Autophagy-Related Proteins; D029701:Saccharomyces cerevisiae Proteins; D006160:Guanosine Triphosphate; D012685:Sepharose; D011494:Protein Kinases; C477823:ATG1 protein, S cerevisiae; C060755:YPT1 protein, S cerevisiae; D020691:rab GTP-Binding Proteins; D005978:Glutathione