title: Freeze-fracture replica immunolabelling reveals human WIPI-1 and WIPI-2 as membrane proteins of autophagosomes.
abstract: Autophagy defines the lifespan of eukaryotic organisms by ensuring cellular survival through regulated bulk clearance of proteins, organelles and membranes. Pathophysiological consequences of improper autophagy give rise to a variety of age-related human diseases such as cancer and neurodegeneration. Rational therapeutic implementation of autophagy modulation remains problematic, as fundamental molecular details such as the generation of autophagosomes, unique double-membrane vesicles formed to permit the process of autophagy, are insufficiently understood. Here, freeze-fracture replica immunolabelling reveals WD-repeat protein interacting with phosphoinositides 1 and 2 (WIPI-1 and WIPI-2) as membrane components of autophagosomes and the plasma membrane (PM). In addition, WIPI-1 is also present in membranes of the endoplasmic reticulum (ER) and WIPI-2 was further detected in membranes close to the Golgi cisternae. Our results identify WIPI-1 and WIPI-2 as novel protein components of autophagosomes, and of membrane sites from which autophagosomes might originate (ER, PM, Golgi area). Hence therapeutic modulation of autophagy could involve approaches that functionally target human WIPI proteins.
authors.affiliation: Autophagy Laboratory, Interfaculty Institute for Cell Biology, Eberhard Karls University Tuebingen, Tübingen, Germany. tassula.proikas-cezanne@uni-tuebingen.de
authors.forename: Tassula
authors.initials: T
authors.lastname: Proikas-Cezanne
authors.forename: Horst
authors.initials: H
authors.lastname: Robenek
mesh_terms: D001343:Autophagy; D000071183:Autophagy-Related Proteins; D002352:Carrier Proteins / Q000378:metabolism* / Q000648:ultrastructure; D045744:Cell Line, Tumor; D005614:Freeze Fracturing / Q000379:methods*; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism* / Q000648:ultrastructure; D010588:Phagosomes / Q000378:metabolism* / Q000648:ultrastructure; D028044:Phosphate-Binding Proteins; D013194:Staining and Labeling
chemical_list: D000071183:Autophagy-Related Proteins; D002352:Carrier Proteins; D008565:Membrane Proteins; D028044:Phosphate-Binding Proteins; C522812:WIPI1 protein, human; C569007:WIPI2 protein, human