title: Proximity Ligation In situ Assay is a Powerful Tool to Monitor Specific ATG Protein Interactions following Autophagy Induction.
abstract: Macroautophagy is a highly regulated intracellular degradation process which has been extensively studied over the last decade. This pathway has been initially described as a non selective process inducing the degradation of parts of the cytoplasm as well as organelles at random. Nevertheless, over the last few years, new research highlighted the existence of a more selective autophagy pathway specifically recruiting some organelles or aggregates to the autophagosomes in order to induce their degradation. These selective autophagy pathways such as aggrephagy, mitophagy, pexophagy or xenophagy, involve the intervention of a cargo, the material to be degraded, cargo adapters, the molecules allowing the recruitment of the cargo to the autophagosome, and the proteins of the ATG8 family which link the cargo adapters to the autophagosome. One of the main questions which now remain is to develop new techniques and protocols able to discriminate between these different types of induced autophagy. In our work, we studied the possibility to use the P-LISA technique, which has been recently developed to study endogenous in vivo protein interactions, as a new technique to characterize the ATG proteins specifically involved in bulk or selective autophagy. In this manuscript, we indeed demonstrate that this technique allows the study of endogenous ATG protein interactions in cells following autophagy induction, but more interestingly that this technique might be used to characterize the ATG proteins involved in selective autophagy.
authors.affiliation: Université de Franche-Comté, Laboratoire de Biochimie, EA3922 « Estrogènes, Expression Génique et Pathologies du Système Nerveux Central », SFR IBCT FED4234, UFR Sciences et Techniques, France.
authors.forename: Thierry
authors.initials: T
authors.lastname: Gauthier
authors.affiliation: Université de Franche-Comté, Laboratoire de Biochimie, EA3922 « Estrogènes, Expression Génique et Pathologies du Système Nerveux Central », SFR IBCT FED4234, UFR Sciences et Techniques, France.
authors.forename: Aurore
authors.initials: A
authors.lastname: Claude-Taupin
authors.affiliation: Université de Franche-Comté, Laboratoire de Biochimie, EA3922 « Estrogènes, Expression Génique et Pathologies du Système Nerveux Central », SFR IBCT FED4234, UFR Sciences et Techniques, France.
authors.forename: Régis
authors.initials: R
authors.lastname: Delage-Mourroux
authors.affiliation: Université de Franche-Comté, Laboratoire de Biochimie, EA3922 « Estrogènes, Expression Génique et Pathologies du Système Nerveux Central », SFR IBCT FED4234, UFR Sciences et Techniques, France.
authors.forename: Michaël
authors.initials: M
authors.lastname: Boyer-Guittaut
authors.affiliation: Université de Franche-Comté, Laboratoire de Biochimie, EA3922 « Estrogènes, Expression Génique et Pathologies du Système Nerveux Central », SFR IBCT FED4234, UFR Sciences et Techniques, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Hervouet
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D051017:Apoptosis Regulatory Proteins; D001343:Autophagy / Q000502:physiology*; D000071190:Autophagy-Related Protein 8 Family; D005455:Fluorescent Antibody Technique; D006801:Humans; D061986:MCF-7 Cells; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D000071456:Sequestosome-1 Protein; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D051017:Apoptosis Regulatory Proteins; D000071190:Autophagy-Related Protein 8 Family; C116838:BNIP3L protein, human; C116741:GABARAP protein, human; C498698:GABARAPL2 protein, human; C502070:MAP1LC3B protein, human; D008565:Membrane Proteins; D008840:Microfilament Proteins; D008869:Microtubule-Associated Proteins; D011518:Proto-Oncogene Proteins; C099718:SQSTM1 protein, human; D000071456:Sequestosome-1 Protein; D025521:Tumor Suppressor Proteins