title: Loss of a membrane trafficking protein αSNAP induces non-canonical autophagy in human epithelia.
abstract: Autophagy is a catabolic process that sequesters intracellular proteins and organelles within membrane vesicles called autophagosomes with their subsequent delivery to lyzosomes for degradation. This process involves multiple fusions of autophagosomal membranes with different vesicular compartments; however, the role of vesicle fusion in autophagosomal biogenesis remains poorly understood. This study addresses the role of a key vesicle fusion regulator, soluble N-ethylmaleimide-sensitive factor attachment protein α (αSNAP), in autophagy. Small interfering RNA-mediated downregulation of αSNAP expression in cultured epithelial cells stimulated the autophagic flux, which was manifested by increased conjugation of microtubule-associated protein light chain 3 (LC3-II) and accumulation of LC3-positive autophagosomes. This enhanced autophagy developed via a non-canonical mechanism that did not require beclin1-p150-dependent nucleation, but involved Atg5 and Atg7-mediated elongation of autophagosomal membranes. Induction of autophagy in αSNAP-depleted cells was accompanied by decreased mTOR signaling but appeared to be independent of αSNAP-binding partners, N-ethylmaleimide-sensitive factor and BNIP1. Loss of αSNAP caused fragmentation of the Golgi and downregulation of the Golgi-specific GTP exchange factors, GBF1, BIG1 and BIG2. Pharmacological disruption of the Golgi and genetic inhibition of GBF1 recreated the effects of αSNAP depletion on the autophagic flux. Our study revealed a novel role for αSNAP as a negative regulator of autophagy that acts by enhancing mTOR signaling and regulating the integrity of the Golgi complex.
authors.affiliation: Department of Human and Molecular Genetics, Virginia Commonwealth University School of Medicine, Richmond, VA, USA.
authors.forename: Nayden G
authors.initials: NG
authors.lastname: Naydenov
authors.forename: Gianni
authors.initials: G
authors.lastname: Harris
authors.forename: Victor
authors.initials: V
authors.lastname: Morales
authors.forename: Andrei I
authors.initials: AI
authors.lastname: Ivanov
mesh_terms: D001343:Autophagy / Q000235:genetics / Q000502:physiology*; D002460:Cell Line; D004848:Epithelium / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism; D006801:Humans; D010588:Phagosomes / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: C119087:ARFGEF1 protein, human; C119088:ARFGEF2 protein, human; C119020:BNIP1 protein, human; C120445:GBF1 protein, human; D020662:Guanine Nucleotide Exchange Factors; D019253:Proto-Oncogene Proteins c-bcl-2; D034741:RNA, Small Interfering; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins