title: LAMP-2 is required for incorporating syntaxin-17 into autophagosomes and for their fusion with lysosomes.
abstract: Autophagy is an evolutionarily conserved process used for removing surplus and damaged proteins and organelles from the cytoplasm. The unwanted material is incorporated into autophagosomes that eventually fuse with lysosomes, leading to the degradation of their cargo. The fusion event is mediated by the interaction between the Qa-SNARE syntaxin-17 (STX17) on autophagosomes and the R-SNARE VAMP8 on lysosomes. Cells deficient in lysosome membrane-associated protein-2 (LAMP-2) have increased numbers of autophagosomes but the underlying mechanism is poorly understood. By transfecting LAMP-2-deficient and LAMP-1/2--double-deficient mouse embryonic fibroblasts (MEFs) with a tandem fluorescent-tagged LC3 we observed a failure of fusion between the autophagosomes and the lysosomes that could be rescued by complementation with LAMP-2A. Although we observed no change in expression and localization of VAMP8, its interacting partner STX17 was absent from autophagosomes of LAMP-2-deficient cells. Thus, LAMP-2 is essential for STX17 expression by the autophagosomes and this absence is sufficient to explain their failure to fuse with lysosomes. The results have clear implications for situations associated with a reduction of LAMP-2 expression.
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Vienna 1090, Austria virginie.hubert@meduniwien.ac.at renate.kain@meduniwien.ac.at.
authors.forename: Virginie
authors.identifier: http://orcid.org/0000-0002-2449-2631
authors.initials: V
authors.lastname: Hubert
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Vienna 1090, Austria.
authors.forename: Andrea
authors.initials: A
authors.lastname: Peschel
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Vienna 1090, Austria.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Langer
authors.affiliation: Core Facilities, Medical University of Vienna, Vienna 1090, Austria.
authors.forename: Marion
authors.initials: M
authors.lastname: Gröger
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Vienna 1090, Austria.
authors.forename: Andrew
authors.initials: A
authors.lastname: Rees
authors.affiliation: Clinical Institute of Pathology, Medical University of Vienna, Vienna 1090, Austria virginie.hubert@meduniwien.ac.at renate.kain@meduniwien.ac.at.
authors.forename: Renate
authors.initials: R
authors.lastname: Kain
keywords: Autophagosome; LAMP-2; LC3; Lysosome; Macroautophagy; STX17; VAMP8